reviewarticle - pdfs.semanticscholar.org€¦ · reviewarticle snake venom pla 2, a promising...

Review ArticleSnake Venom PLA2 a Promising Target for Broad-SpectrumAntivenom Drug Development

Huixiang Xiao Hong Pan Keren Liao Mengxue Yang and Chunhong Huang

Department of Biochemistry College of Basic Medical Sciences Nanchang University Nanchang Jiangxi Province China

Correspondence should be addressed to Chunhong Huang chhuangncueducn

Huixiang Xiao and Hong Pan contributed equally to this work

Received 5 September 2017 Accepted 30 October 2017 Published 29 November 2017

Academic Editor Ji-Fu Wei

Copyright copy 2017 Huixiang Xiao et al This is an open access article distributed under the Creative Commons Attribution Licensewhich permits unrestricted use distribution and reproduction in any medium provided the original work is properly cited

Snakebite envenomation is a neglected global health problem causing substantial mortality disability and psychological morbidityespecially in rural tropical and subtropical zones Antivenin is currently the only specificmedicine for envenomation However it isrestricted by cold storage snakebite diagnosis and high price Snake venom phospholipase A2s (svPLA2s) are found in all kinds ofvenomous snake families (eg Viperidae Elapidae andColubridae) Alongwith their catalytic activity svPLA2s elicit a wide varietyof pharmacological effects that play a pivotal role in envenomation damage Hence neutralization of the svPLA2s could weakenor inhibit toxic damage Here we overviewed the latest knowledge on the distribution pathophysiological effects and inhibitors ofsvPLA2s to elucidate the potential for a novel wide spectrum antivenom drug targeting svPLA2s

1 Introduction

Snakebite envenomation is a critical public health problemand fieldwork hazard causing high mortality and mor-bidity particularly in tropical and subtropical regions Asmost ophidian incidents occur in rural areas of developingcountries accurate statistical data concerning the numberof victims is difficult to obtain [1] As extrapolated byChippaux worldwide 5400000 people are bitten by snakes2500000 are envenomed 125000 die and more than100000 individuals suffer from severe sequelae each year [2]Unfortunately snakebite was neglected by governments andinternational health agencies for a long time even thoughthe snake bite mortality rate is equivalent to one-fifth ofthe deaths from malaria worldwide and half of the deathsfrom HIVAIDS in India [3] In 2009 the World HealthOrganization (WHO) recognized snake bite as a neglectedtropical disease [1] Currently antivenin is the only specifictreatment towards envenomation Although the immunizedanimal sera (mainly horse or sheep) presently used are highlyeffective they are limited by a few drawbacks [4] First localtissue damage resulting from snake venom exposure oftenleading to amputation cannot be reversed by antivenin [4]Furthermore early and late adverse reactions to antivenin

(eg anaphylaxis pyrogenic reactions and serum sickness)occur in some cases [5] Additionally access to antiveninsis often limited Some remote rural communities whereantivenoms are most needed cannot get adequate suppliesdue to the lack of cold chain storage and other complexpolitical reasons Finally most antivenoms are too expensivefor the patientrsquos family in low-income countries [6]

Recently the nonprofit French drug firm Sanofi Pasteurhad ceased the production of Fav-Afrique the most effectiveantivenin against Africarsquos vipers mambas and cobras Thishas resulted in a large-scale snakebite crisis in rural Africa [7]This alarming situation demonstrates the need for antiveninreplacements and new antivenom drug candidates Thisreview article focuses on snake venom phospholipase A2s(svPLA2s) a chemical family that is widely distributed invenomous snake species Here we describe svPLA2s theantienvenomation effects of their inhibitors and the potentialof being a common target for broad-spectrum antivenomdrugs

2 Characteristics of svPLA2

Snake venoms are complicated mixtures consisting of phos-pholipase A2s metalloproteases C-lectins serine proteases

HindawiBioMed Research InternationalVolume 2017 Article ID 6592820 10 pageshttpsdoiorg10115520176592820

2 BioMed Research International

L-amino acid oxidases disintegrins and a few other com-pounds [1] Most svPLA2s hydrolyze glycerophospholipids atthe sn-2 position of the glycerol backbone freeing lysophos-pholipids and fatty acids svPLA2s share 44ndash99 aminoacid identity in their primarily structure which results tohigh similarity in their tertiary structure [8] Based on theirsize location function substrate specificity and calciumrequirement PLA2s are classified into six families svPLA2belongs to the secretory PLA2 (sPLA2) family (groups IA IIAand IIB) [9ndash11] Cobras and kraits rattlesnakes and Gaboonvipers have svPLA2s in groups IA IIA and IIB respectively[8] There are also group IB enzymes which are mainlyfound in mammalian pancreas that have been reported insome snake venoms such as Oxyuranus scutellatus [12]Pseudonaja textilis [13] and Micrurus frontalis frontalis [14]These compounds are conserved in structure andhave similarmolecular masses (sim10ndash20 kDa) 5ndash7 disulfide bonds andanalogous three-dimensional structures [15] InGroup I thereare approximately 115ndash120 residues 7 disulfide bonds (theunique disulfide linking residues 11 and 77) and G IA has acharacteristic surface loop between residues 63 to 67 calledelapidic loop [11] While G IB has a five amino acids residues(residues 62ndash67) extension termed pancreatic loop some GIB snake venom PLA2 even has an eight-residue propeptidesegment in their mature state [13 16] In contrast GroupII has a C-terminal extension the unique disulfide linkingresidues 50 and 137 GIIA have a 7-residue C-terminal exten-sion and seven conserved disulfide bonds while inGroup IIBthe C-terminal extension is 6 residues and only six disulfidesremained in which a universally conserved 61ndash95 disulfideis lacking [11] Furthermore a new subgroup (Lys49 PLA2homologues) can be created through mutation Replacementof the 49th residue (asparagine) with lysine results in aninactive or weakly toxic PLA2 This lysine residue can alsointeractwith other amino acids in the ldquocalcium-binding looprdquoresulting in the loss of calcium-dependent catalytic activity[17 18] Most svPLA2s exist as monomers but some exist incomplexes which mainly exhibit presynaptic neurotoxicitythrough combination of isoenzymes or other proteins [19]

3 PLA2s Are Extensively Distributed inSnake Venom

Mackessy [20] analyzed crude venom from the main cladesof venomous snakes via SDS-PAGE and found that svPLA2sexisted in almost every family (Figure 1) The highestamounts were found in Elapidae Viperidae and Hydrophi-idae The lowest were found in Colubridae (which is usuallynonvenomous) Through the application of transcriptomicsand proteomics we gained a better understanding of venomcomposition and the pharmacological properties of thevenom components [21] Betzel et al found that PLA2smade up 32ndash598 in Viperidae snake venom [22] HoweverBungarus fasciatus venom was found to consist of up to 71of PLA2s [23] Moreover Gutierrez and Lomonte found thatthemost lethal fractions inMicrurus fulvius (family Elapidae)were two PLA2 molecules which represented 334 of thewhole venom [24] To date more than 464 unique svPLA2shave been recorded in UniProtKB database What has been

presented above indicates that PLA2s are abundant and fataltoxins in most snake venoms

4 svPLA2s Have a Wide Spectrum ofPharmacological Effects

Despite producing lysophospholipids and fatty acid proin-flammatorymediators svPLA2s also present a wide spectrumof pharmacological effects in victims (ie neurotoxicitymyotoxicity anticoagulant effects cytotoxicity cardiotoxic-ity and edema Table 1) The diverse toxic effects are tightlyrelated to the multiple functional sites on the surface ofsvPLA2s and their different binding receptors [25]

41 svPLA2 Neurotoxicity Neurotoxic svPLA2s can blockneuromuscular transmission in vertebrate skeletal musclescausing acute neuromuscular weakness and paralysis result-ing in respiratory depression and death [53] NeurotoxicsPLA2s are mainly found in the Elapidae (kraits elapidsand coral snakes) and Viperidae (vipers and rattlesnakes)Their toxicity varies greatly among species ranging from1 120583gkg (Textilotoxin) to 380120583gkg (HDP-2 from Viperanikolskii) [53] Previous studies indicate that there is nocorrelation between toxicity and PLA2 hydrolysis activitysvPLA2 neurotoxicity affects presynaptic nerve terminalsso these compounds are commonly referred as presynap-tic neurotoxins or 120573-neurotoxins (120573-ntxs) [54] 120573-ntxs aremonomers or noncovalent complexes containing 2ndash5 sub-units with at least one PLA2 subunit To our knowledge all 120573-ntxs hydrolyze phospholipids especially anionic lipids (egphosphatidylserine phosphatidic acid and phosphorylatedphosphatidylinositols) which are abundant in the cytosolicleaflets of organelles and the plasmamembrane of eukaryoticcells [55] svPLA2s also bind to special tissue sites to achievetheir neurotoxicity effects The mechanism of svPLA2 neuro-toxicity is still under investigation

42 svPLA2 Myotoxicity svPLA2s can induce acute necrosisof skeletal muscle (myonecrosis) [56] In the envenomationthismyonecrosis can potentially lead to permanent tissue lossor amputation [57] svPLA2 myotoxins are mainly found invenom from Elapidae including sea snakes and Viperidae[58] Depending on the venom these svPLA2s can elicitlocal or systemic myotoxicity Local myotoxicity is mainlyelicited by viperid venom This damage is limited to theregion where the toxin is injected and is often coupledwith hemorrhaging blistering and edema [57 59] Systemicmyotoxicity is elicited by elapid venom (ie some sea snaketerrestrial elapids) This causes muscle damage and a distinctincrease of creatine kinase (CK) activity in plasma and isassociated with renal failure and myoglobinuria [58] Alongwith sharing a highly conserved structure svPLA2myotoxinsare tightly associatedwith neurotoxins Both achieve a similarcellular lesion through membrane perturbation cytosolicCa2+ homoeostasis imbalance and cell degeneration [60]Furthermore some neurotoxic svPLA2s (eg notexin andcrotoxin) cause acute skeletal muscle necrosis adding tosystemic toxic effects (ie rhabdomyolysis) [60]

BioMed Research International 3

M (kD)N a

ter

P po

rphy

riacu

s

A a

ntar

cticu

s

H c

yano

cinct

usL

har

dwick

iiL

sem

ifasc

iata

N n

naj

a

N m

elano

leuca

N n

igric

ollis

M f

ulvi

us

Stan

dard

s

B ir

regu

laris

B d

endr

ophi

la

A n

asut

a

T b

iscut

atus

lam

bda

A p

orto

ricen

sis

L m

adag

asca

riens

is

Typical protein familyactivity

Acetylcholinesterasemetalloprotease

Serine protease()

CRiSP

Phospholipase A2

ree-nger toxins

2000

974

664

556

366

310

210

142

60

35

(a)

T b

orne

ensis

T p

unice

us

T st

ejneg

eri

T fl

avov

iridi

sT

pur

pure

omac

ulat

us

D ru

sselli

i rus

selli

i

C rh

odos

tom

a

A n

itsch

iiC

resim

us

E ca

rinat

us so

chur

eki

B ga

boni

ca ga

boni

ca

B a

rieta

ns

V ra

ddei

V a

mm

odyt

es

C ti

gris

C tr

ansv

ersu

s

stan

dard

s

M (kD)Typical protein familyactivity

NucleasesL-amino acid oxidaseMetalloprotease-PIII

Serine proteases

Metalloprotease-PI

PLA2C-type lectin

CRiSP

Disintegrins

Myotoxins()

2000

974

664

556

366

310

210

142

60

35

(b)

Figure 1 SDS-PAGE profile of major venom components in the main clades of venomous snakes (adapted from [20]) (a) Families Elapidaesubfamilies Elapinae Laticaudinae Hydrophiinae and Colubrinae (b) Family Viperidae and subfamilies Crotalinae (C) and Viperinae (V)Ovals enclose some bands that are typical of protein families based on published mass () indicates hypothetical protein family or activity

Residue 49 in myotoxic svPLA2s is usually associatedwith PLA2 enzymatic activity Asp49-PLA2s are generallystrongly catalytic whereas Lys49 homologues are either notcatalytic or weakly catalytic There are also other aminoacid substitutions such as Ser49 Arg49 Asn49 or Gln49

[56] The lysophospholipids released from phospholipid thathydrolyzed by Asp49 PLA2 usually cause skeletal musclenecrosis via direct disruption of membrane stabilizationandor indirect biophysical alteration of membrane [61] TheLys49PLA2myotoxins are devoid of catalytic activity existing


Table1Featurestoxicitiesbinding

receptorsandenzymaticactiv

ityof

snakev

enom

PLA2s

Nam

eSn

akes

pecies

Structuralfeatures

subtyp

eaTo

xicitie

sLethality

inmou

se(120583gkg)b

Bind

ingproteins

intissuec

PLA2activ

ity(120583molm

inm

gtoxin)

dRe

ference

Neurotoxin

Crotoxin

Crotalus

durissusterrifi

cus

Heterod

imericA

IIA-sPL

A2-like

Neurotoxicitym

yotoxicitycardiotoxicity

60ndash240

(iv)

Crocalbin

CaM

85[26]

BIIA

-sPL

A2

MsPLA2-I

Micr

urus

spixii

Mon

omericIA-

PLA2

Neurotoxicitym

yotoxicityantiplasm

odialactivity

edem

and

nAchR

Yes

[27]

Taipoxin

Oxyuranus

scutellatus

Trim

eric120572

IAtoxic120573

IA-

sPLA2lik

e120574IB-sPLA2

glycosylated

Presyn

aptic

neurotoxicity

2(iv)

M-sPL

A2R

NP

TCBP

-49

04

[28ndash30]

cytotoxicity

Textilo

toxin

Pseudona

jatextilis

Pentam

ericA

BandCareIA-

sPLA2D2

identicalS-SlinkedIB-sPLA2sglycosylated

Presyn

aptic

neurotoxicity

1(iv)

M-sPL

A2R

32

[132831]

Ammod

ytoxin

Vipera

ammodytes

Mon

omericIIA-sPL

A2

Presyn

aptic

neurotoxicity

21

(iv)

M-sPL

A2R

CaMP

DI

FXa14-3-3

proteins

280

[32ndash35]

antic

oagu

lant

120573-Bun

garotoxin

Bungarus

multicinctus

Dim

ericA

IA-

sPLA2

Presyn

aptic

neurotoxicity

19ndash130

(ip)

v-d

K+channel

61[3637]

S-Slin

kedto

subu

nitB

BPT

I-lik

e

Notexin

Notechisscutatus

Mon

omericIA-sPL

A2(A

sp49)

Myotoxicitypresynapticneurotoxicity

neph

rotoxicity

17(iv)

nd

1390

[3839]

Myotoxin

MyotoxinIII

Bothrops

asper

Dim

ericIIA

-sPL

A2(A

sp49)

Myotoxicity

470(iv)

nd

750

[40]

antic

oagu

lantedema

MyotoxinII

Bmoojen

iMon

omericIIA-sPL

A2(Lys49)

Myotoxicityedema

7600

(ip)

nd

Non

e[41]

CoaTx

-II

Crotalus

oreganus

abyssus

Dim

ericIIA-sPL

A2(Lys49)

Myotoxicityedemaantib

acteria

lactivity

nd

nd

Non

e[42]

Cr5

Calloselasm

arhodostoma

Mon

omericIIA-sPL

A2(Lys49)

Cytotoxicitym

yotoxicityedema

70(icv

)nd

Non

e[43]

BaTX

Bothrops

alternatus

Mon

omericIIA

-sPL

A2(Lys49)

Cytotoxicitym

yotoxicityedemaneurotoxicity

7000

(iv)

nd

Non

e[44]

Cr-IV1

Calloselasm

arhodostoma

Mon

omericIIA-sPL

A2(A

sp49)

Myotoxicitycytotoxicity

edema

70(icv

)nd

0014

[45]

Ammod

ytin

LVipera

ammodytes

Mon

omericIIA-sPL

A2(Ser49)

Myotoxicity

3600

(ip)

nd

Non

e[46]

Anticoagulant

Daboxin

PDaboiarusse

liiMon

omericIA-

sPLA2

Strong

antic

oagu

lant

nd

FXFXa

1140

[47]

RVV-PFIIc1015840

Drusselii

Mon

omericIIA-sPL

A2(A

sp49)

Anticoagu

lant

100(ip)

nd

Yes

[48]

CM-IV

Najanigrico

llis

Mon

omericIIA-sPL

A2(A

sp49)

Strong

lyantic

oagu

lantpresynapticneurotoxicity

180(ip)

FXaFV

IIaYes

[4950]

CM-II

Najamossambica

Mon

omericIA-

sPLA2

Weakanticoagu

lantm

yotoxicityn

eurotoxicity

nd

TFFVII

Yes

[5152]

a BPT

Ibo

vine

pancreatictrypsin

inhibitor

b icv

intracerebroventric

ularivintraveno

usicintraciste

rnalipintraperito

nealn

dno

tdeterminedcCa

Mcalmod

ulinN

Pneuron

alpentraxin

PDIprotein

disulfide

isomeraseTC

BP-49taipoxin-associatedcalcium-binding

protein49M

-sPL

A2R

M-ty

pesPLA2receptorFxablood

coagulationfactor

XaFX

bloo

dcoagulationfactor

XTF

tissue

factorFVIIblood

coagulationfactor

VIIFVIIa

blood

coagulationfactor

VIIa

v-d

K+channelvoltage-dependent

K+channels

d pho

spho

lipaseA2activ

ityisin120583molm

inm

gof

toxin

Yesoriginalresearch

paperd

oesn

otshow

phosph

olipaseA2activ

ityin

concretenu

mbero

rnot

in120583molm

inm

gof

toxin

Non

eallP

LA2ho

mologuesa

rehere

considered

tobe

enzymaticallyinactiv

eAd

aptedfro

m[5051]


as homodimers in solution connected by noncovalent bonds[56] Previous studies focused on the fact that amino acidscomposition of synthetic peptides has revealed that the C-terminal regions of 115ndash129 residues which are positivelycharged and full of basic aromatic hydrophobic residues arethe key structure in eliciting myotoxic effects [62 63] Site-directedmutagenesis experiments proved that Tyr117 Arg118Tyr119 Lys122 and Phe125 also have significant impacts onmyotoxicity [64]

43 svPLA2 Anticoagulant Effect The anticoagulant effect ofsvPLA2 usually causes bleeding in victimprey by inhibitingone or two steps in the blood coagulation cascade PLA2scan be classified as strong weak and nonanticoagulant basedon the dose required to inhibit blood coagulation [65] Thehydrolysis of phospholipids by svPLA2 would be the pri-mary mechanism to account for PLA2srsquo anticoagulation [66]However in the absence of phospholipids some svPLA2scould also inhibit coagulation [67] The correlation betweensvPLA2 enzymatic activity and anticoagulant effect is stillunknown Furthermore there are other mechanisms thatrestrain coagulation such as inhibition of the activation of theconversion of FX (blood coagulation factor X) to FXa (bloodcoagulation factor Xa) andor prothrombin to thrombin [68]

svPLA2s can also induce other toxic effects such asmyoglobinuria-inducing hemolytic and platelet aggregationinitiatinginhibiting activities [49] Their wide distributionconserved structures and various severe pharmacologicaleffects suggest that svPLA2s represent a promising target fornew antivenommedicine Indeed there is sufficient evidencethat PLA2 inhibitors (PLIs) are effective in using snake venomenvenomation therapy [69]

5 PLA2 Inhibitors Attenuate Morbidity andMortality of Snakebite Envenomation

Due to the high cost long production period limited cat-egories short storage life and common clinical side-effectsof current antivenin scientists have attempted to create anti-dotes from herbal extracts marine compounds mammalianand snake serum and modified chemical molecules andpeptides [70] svPLA2s are the ideal target and widely usedfor antidote screening Indeed both natural and syntheticsvPLA2 inhibitors are able to attenuate the morbidity andmortality of snakebite envenomation

51 Natural svPLA2 Inhibitors from Plants Marine Extractsand Mammalian Serum Medicinal plant extracts as tradi-tional antidotes have long been used in countries where theurotherapy is unobtainable [71] In addition these traditionaland herbal treatments are often used as adjuvant therapiesalong with the antivenin treatment Most plant antitoxicagents function by neutralizing svPLA2rsquos toxicity An activeglycoprotein (WSG) from Withania somnifera completelyinhibits the cytotoxicity edema and myotoxicity of NN-Xia-PLA2 isolated from Naja atra venom but fails to neutralizethe neurotoxicity [72ndash74] WSG has a similar structure to the120572-chain of the PLIs derived from Australian elapid serum

and was found to interact with NN-XIandashPLA2 but themechanism currently remains unknown [74]

The aqueous extract of Casearia sylvestris was foundto be effective against two snake venom toxins (Asp49-PLA2 and Lys49-PLA2 isolated from venom of B moojeniB pirajai B neuwiedi and B jararacussu) Indeed thisplant has been found to inhibit myotoxicity hemorrhageanticoagulation and edema [75 76] It is also able to preventmyonecrosis initiated by two Lys49-PLA2 toxins (PrTX-Ifrom B pirajai and BthTX-I from B jararacussu venom)and neuromuscular blockages [77] Recently research hasshown that human secretory PLA2 inhibitors (eg quercetinbiflavonoid morelloflavone [78 79]) isolated from plantextracts can also inhibit svPLA2

Marine organisms are also a reservoir for antivenomsManoalide (MLD) a natural product from sponge Luffariellavariabilis can irreversibly inhibit extracellular PLA2 activityof cobra and rattlesnake venom with an IC50 value of 19 and07 120583M respectively [80] Its synthetic analogue manoalogue(MLG) is also inhibitive to cobra PLA2 activity with an IC50value of 75120583M [81]

Natural svPLA2 inhibitors also exist in some mammalianserums DM64 is an acidic glycoprotein isolated from serumof the opossumDidelphis marsupialis DM64 can completelyprevent myofiber breakdown caused by myotoxins I (Asp49)and II (Lys49) of B asper venom [82] N-glycosylation sites(Asn46 Asn179 Asn183 and Asn379) in this antimyotoxicprotein play important roles in this inhibitory action [83]

52 Snake Blood PLA2 Inhibitors Many venomous and non-venomous snake species are naturally resistant to the deleteri-ous actions of snake venom components In many cases thisis due to the presence of specific antitoxins circulating in theirblood These alexeteric factors are proteins generated in thesnakersquos liver with native molecular masses ranging from 75to 180 kDa These nonimmunoglobulin antitoxins are PLA2inhibitors (ie snake blood phospholipase A2 inhibitorssbPLIs) and are used to protect the snake from the internalor external envenomation

sbPLIs can be produced by snakes of the ElapidaeViperidae Hydrophidae Colubridae and Boidae familiesThese sbPLIs can be classified into three groups based onthe homology of their amino acid sequence 120572 120573 and 120574 [84]Generally the 120572 and 120574 sbPLIs simultaneously occur in severalsnake species while the 120573sbPLIs have only been reportedin three snake species When the target PLA2s are Lys49homologues or Asp49 myotoxins the sbPLIs are specificallycalled myotoxin inhibitor proteins (MIPs) [85 86]

Since the first 120572PLI (BaMIP) was isolated from B asperserum 15 kinds of 120572sbPLIs have been discovered in the dif-ferent venomous snake families Previous studies have shownthat BaMIP can block bothmyotoxins I and III (isolated fromB asper venom) [87] The 120572PLIs 120572TfPLI and 120572AbsPLI alsoshow good inhibition of the enzymatic activities of acid-PLA2(isolated from Viperidae) CgMIP-II and AnMIP can inhibitthe basic-PLA2 enzymatic activities of Viperidae venomBaMIP BmjMIP and BjussuMIP can inhibit the enzymaticactivities and toxic effects (ie edema myotoxicity andcytotoxicity) of acidbasic-PLA2 Furthermore Quiros et al


extracted a new myotoxin inhibitor 120572PLI from A nummiferserum (AnMIP) and found that this protein at a ratio of 1 1could decrease 67 of the A nummifer myotoxin II and 93of the B asper myotoxin I [85]

Currently four kinds of 120573sbPLIs have been found inthree snake species 120573 PLI specifically inhibits the basic-PLA2 enzymatic activities of Viperidae The first 120573sbPLI waspurified from G brevicaudus as a homotrimer and is specificfor basic-PLA2s from homologous venoms and forms a stablePLA2-120573sbPLI complex at a molar ratio of 1 1 [88]

Twenty-three types of 120574sbPLIs have been found in ven-omous and nonvenomous species 120574PLI from Elapidae andother nonvenomous snakes can inhibit PLA2 activity in arange of different snake venoms We recently reported anovel 120574PLI isolated from the serum of Sinonatrix annularisnamed 120574saPLI that showed a strong inhibition of lecithindegradation elicited by D acutus venom PLA2s in an in vitrostudy [89] The 120574saPLI was also effective in the inhibition ofhemorrhagic toxicities elicited by D acutus N atra and Ahalys venom [90]

53 Poly or Monoclonal Antibodies of svPLA2 Are Effective inNeutralizing Snake Venom Unlike the common antiveninsof venom proteome Garcia Denegri et al developed a poly-antibody using a nontoxic PLA2 (BaSpII RP4) from Bothropsalternatus as antigen [91]This antibody showed a specific andsensitive inhibition of the venom PLA2srsquo enzymatic activityFurthermore the myotoxicity and mortality of the crudevenom were significantly reduced in the presence of anti-PLA2 IgG When treated with a high dose of 2 times LD50equivalent to 112 120583g of B alternatus venom and 262mg ofIgG all of the test animals survived after 48 h In contrastthe control group (112 120583g venom preincubated with PBS)died within 4 hours 525mg of IgG treated animals couldeven endure as high as 4 times the LD50 dose of venom(224120583g) with half of the treated group remaining alive atthe end of 48 h In contrast the control group (224120583g venompreincubated with PBS) died shortly within 90mins

Rodriguez et al also produced a IgG against crotoxin(a basic PLA2) the principle toxin of C durissus terrifi-cus (Cdt) with high myotoxic and neurotoxic activitiesMice preincubated with the anticrotoxin IgG showed lowmortality after 24 and 48 h of inoculation (at 4120583g Cdtvenomtest animal) The investigation showed that the IgGsof anti-PLA2 were more effective than anticrotalic serumat neutralizing lethal activity [92] Additionally the anti-PLA2 IgGs raised via immunization with P9a or P10a twotypes of less toxic Cdt-PLA2s cross-reacted with all theisoforms of PLA2s in the Cdt venom [93] Although theseantitoxic effects were only tested with their original venomsthe wide cross-reaction of these anti-PLA2 IgGs with othersvPLA2s suggested that these compounds could likely also beused to neutralize other snake venoms In other words theimproved neutralization activity of these anti-svPLA2 IgGsindicates svPLA2s are a promising target for broad-spectrumantivenom drug development

54 Artificial Inhibitor of Mammal PLA2 Exhibits EffectiveAntivenom Activity Varespladib (LY315920) was designed as

an inhibitor of the IIa V and X isoforms of the mammaliansecretory phospholipase A2 (sPLA2) This compound actsas an anti-inflammatory agent by disrupting the first stepof the arachidonic acid pathway of inflammation From2006 to 2012 varespladib was under active investigation byAnthera Pharmaceuticals for using as a potential therapyfor several inflammatory diseases including acute coronarysyndrome and acute chest syndrome [94 95] Thought tobe an effective antiatherosclerotic agent varespladib showedpromising therapeutic effects in reducing plasma sPLA2 andlow-density lipoprotein (LDL) [96]

Varespladib has recently been repurposed as an effectivebroad-spectrum svPLA2 inhibitor and used for treatment ofsnakebite envenomation Varespladib and its orally bioavail-able prodrug methyl-varespladib (LY333013) showed stronginhibitory ability of 28 kinds of svPLA2s from six continentsIndeed the IC50 values ranged from nano- to picomolarsin an in vitro experiment [97] Additionally the compoundelicited surprising effects with eastern coral snake (Micrurusfulvius) venom which was considered to have the highestsPLA2 activity and most intense hemo- and neurotoxiceffects Pretreatment with 01mg of varespladib prolongedsurvival in mice at 4 times the LD50 dose of eastern coralsnake venom over the course of 8 h All the negative controlmice died at an average of 63min whereas the varespladibtreatment group survived for an average of 1140min Vare-spladib also showed promising in vivo protection in Viperaberus envenomed mice Mice treated with a subcutaneousinjection of a 100 lethal dose of venom and varespladibsurvived for more than 24 h [97] These findings are solidevidence of svPLA2 being the target for a broad-spectrumantivenom

6 Conclusions

svPLA2s are widely distributed in snake venoms A svPLA2could elicit one or more pharmacological effects (eg neu-rotoxicity myotoxicity anticoagulant and edema) Further-more svPLA2s can interact with other svPLA2s (eg twodifferent svPLA2s the ldquoAsprdquo and ldquoLysrdquo myotoxins fromBothrops asper have been shown to synergistically enhancemyonecrosis in in vitro and in vivo studies [98]) or othervenom components (eg taicatoxin a Ca2+ channel inhibitorcomposed of an 120572-neurotoxin-like peptide a neurotoxicphospholipase A2 and a serine protease inhibitor connectedby noncovalent bonds [99])

A variety of PLA2 inhibitors were discovered or syn-thesized in the past few decades Most inhibitors extractedfrom medical plants marine animals and mammalianserum specially inhibit svPLA2 toxicity sbPLIs are naturalendogenous protective components against snake venomamong which the 120574PLI were commonly inhibitive to differ-ent category of venoms [100] Anti-PLA2 antibodies couldspecifically inactivate enzymatic activity and toxicity bothwith the original venom and other svPLA2s [93] Indeedsome of these compounds could function even better thanthe antivenin that is currently clinically applied [92] Asynthetic human sPLA2 inhibitor varespladib was found topossess the ability to neutralize a variety of snake venoms


worldwide with significant prolongation of survival time onrats that were inoculated with varespladib simultaneouslyor following exposure [97] In conclusion the anti-PLA2drugs are promising antidotes for a broad-spectrum of snakevenoms and other animal toxins and could also be effectivein prevention of inflammatory reactions (ie systemic toxi-cological syndromes)

Conflicts of Interest

The authors confirm that this article content has no conflictsof interest

Authorsrsquo Contributions

HuixiangXiao andHongPan contributed equally to thisworkand are considered as co-first authors

Acknowledgments

The authors are grateful for the support of the NationalNatural Science Foundation of China (no 31260209 and no31460227) Natural Science Foundation of Jiangxi Province(20171BAB204015) and Cultivating Foundation of YoungScientists of Jiangxi Province (20171BCB23018)

References

[1] D AWarrell ldquoSnake biterdquoTheLancet vol 375 no 9708 pp 77ndash88 2010

[2] J-P Chippaux ldquoSnake-bites appraisal of the global situationrdquoBulletin of theWorld Health Organization vol 76 no 5 pp 515ndash524 1998

[3] Editorial ldquoSnake bitemdashthe neglected tropical diseaserdquo Lancetvol 386 no 9999 pp 1110 2015

[4] J M Gutierrez R D G Theakston and D A Warrell ldquoCon-fronting the neglected problem of snake bite envenoming theneed for a global partnershiprdquo PLoS Medicine vol 3 no 6 pp0727ndash0731 2006

[5] HADe SilvaNMRyan andH J De Silva ldquoAdverse reactionsto snake antivenom and their prevention and treatmentrdquoBritish Journal of Clinical Pharmacology vol 81 no 3 pp 446ndash452 2016

[6] J M Gutierrez D Williams H W Fan and D A WarrellldquoSnakebite envenoming from a global perspective towards anintegrated approachrdquoToxicon vol 56 no 7 pp 1223ndash1235 2010

[7] Q Schiermeier ldquoAfrica braced for snakebite crisisrdquo Nature vol525 no 7569 p 299 2015

[8] D L Scott ldquoPhospholipase A2 structure and catalytic proper-tiesrdquo in In Venom Phospholipase A2 Enzymes Structure Func-tion and Mechanism R M Kini Ed pp 97ndash128 John WileyChichester UK 1997

[9] E A Dennis J Cao Y-H Hsu V Magrioti and G KokotosldquoPhospholipaseA2 enzymes physical structure biological func-tion disease implication chemical inhibition and therapeuticinterventionrdquo Chemical Reviews vol 111 no 10 pp 6130ndash61852011

[10] R H Schaloske and E A Dennis ldquoThe phospholipase A2superfamily and its group numbering systemrdquo Biochimica etBiophysica Acta (BBA) - Molecular and Cell Biology of Lipidsvol 1761 no 11 pp 1246ndash1259 2006

[11] D A Six and E A Dennis ldquoThe expanding superfamily ofphospholipase A2 enzymes classification and characterizationrdquoBiochimica et Biophysica Acta (BBA) - Molecular and Cell Biol-ogy of Lipids vol 1488 no 1-2 pp 1ndash19 2000

[12] J Fohlman P Lind and D Eaker ldquoTaipoxin an extremelypotent presynaptic snake venom neurotoxin Elucidation ofthe primary structure of the acidic carbohydrate-containingtaipoxin-subunit a prophospholipase homologrdquo FEBS Lettersvol 84 no 2 pp 367ndash371 1977

[13] J A Pearson M I Tyler K V Retson and M E H HowdenldquoStudies on the subunit structure of textilotoxin a potent presy-naptic neurotoxin from the venom of the Australian commonbrown snake (Pseudonaja textilis) 3 The complete amino-acidsequences of all the subunitsrdquo Biochimica et Biophysica Acta(BBA) - Protein Structure and Molecular Enzymology vol 1161no 2-3 pp 223ndash229 1993

[14] B R Francis N Jorge Da Silva Jr C Seebart L L Casais ESilva J J Schmidt and I I Kaiser ldquoToxins isolated from thevenomof the Brazilian coral snake (Micrurus frontalis frontalis)include hemorrhagic type phospholipases A2 and postsynapticneurotoxinsrdquo Toxicon vol 35 no 8 pp 1193ndash1203 1997

[15] R C De Paula H C Castro C R Rodrigues P A Meloand A L Fuly ldquoStructural and pharmacological features ofphospholipases A2 from snake venomsrdquo Protein and PeptideLetters vol 16 no 8 pp 899ndash907 2009

[16] S P Mackessy ldquoSnake Venom Phospholipase A2 Enzymesrdquo inHandbook of Venoms and Toxins of Reptiles S P Mackessy Edpp 174ndash195 Taylor and Francis Boca Raton Fla USA 2010

[17] T Petan I Krizaj and J Pungercar ldquoRestoration of enzymaticactivity in a Ser-49 phospholipase A2 homologue decreases itsCa2+-independent membrane-damaging activity and increasesits toxicityrdquo Biochemistry vol 46 no 44 pp 12795ndash12809 2007

[18] R J Ward L Chioato A H C De Oliveira R Ruller and JM Sa ldquoActive-site mutagenesis of a Lys49-phospholipase A2Biological andmembrane-disrupting activities in the absence ofcatalysisrdquo Biochemical Journal vol 362 no 1 pp 89ndash96 2002

[19] C Bon ldquoMulticomponent neurotoxic phospholipases A2rdquo inVenom Phospholipase A2 Enzymes Structure Function andMechanism R M Kini Ed pp 269ndash285 John Wiley Chich-ester UK 1997

[20] S P Mackessy ldquoThe field of reptile toxinology snakes lizardsand their venomsrdquo in In Handbook of Venoms and Toxins ofReptiles S P Mackessy Ed pp 3ndash19 Taylor and Francis BocaRaton Fla USA 2010

[21] J J Calvete ldquoProteomics in venom research a focus on PLA2moleculesrdquo Acta Chimica Slovenica vol 58 no 4 pp 629ndash6372011

[22] D Georgieva R K Arni and C Betzel ldquoProteome analysis ofsnake venom toxins pharmacological insightsrdquo Expert Reviewof Proteomics vol 5 no 6 pp 787ndash797 2008

[23] R H Ziganshin S I Kovalchuk G P Arapidi et al ldquoQuan-titative proteomic analysis of vietnamese krait venoms neuro-toxins are the major components in bungarus multicinctus andphospholipases A2 in bungarus fasciatusrdquo Toxicon vol 107 pp197ndash209 2015

[24] J M Gutierrez and B Lomonte ldquoPhospholipases A2 unveilingthe secrets of a functionally versatile group of snake venomtoxinsrdquo Toxicon vol 62 pp 27ndash39 2013

[25] R M Kini ldquoExcitement ahead structure function and mecha-nism of snake venom phospholipase A2 enzymesrdquo Toxicon vol42 no 8 pp 827ndash840 2003


[26] S C Sampaio S HyslopM RM Fontes et al ldquoCrotoxin novelactivities for a classic 120573-neurotoxinrdquo Toxicon vol 55 no 6 pp1045ndash1060 2010

[27] A L C Terra L S Moreira-Dill R Simoes-Silva et al ldquoBiolog-ical characterization of the amazon coral micrurus spixii snakevenom isolation of a new neurotoxic phospholipase A2rdquo Toxi-con vol 103 pp 1ndash11 2015

[28] G Lambeau P Ancian J Barhanin and M LazdunskildquoCloning and expression of a membrane receptor for secretoryphospholipases A2rdquo The Journal of Biological Chemistry vol269 no 3 pp 1575ndash1578 1994

[29] G Lambeau A Schmid-AllianaM Lazdunski and J BarhaninldquoIdentification and purification of a very high affinity bindingprotein for toxic phospholipases A2 in skeletal musclerdquo TheJournal of Biological Chemistry vol 265 no 16 pp 9526ndash95321990

[30] B V Lipps ldquoIsolation of subunits 120572 120573 and 120574 of the complextaipoxin from the venom of Australian taipan snake (Oxyu-ranus s scutellatus) characterization of 120573 taipoxin as a potentmitogenrdquo Toxicon vol 38 no 12 pp 1845ndash1854 2000

[31] A Coulter R Harris A Broad et al ldquoThe isolation and someproperties of the major neurotoxic component from the venomof the common or Eastern Australian brown snake (Pseudonajatextilis)rdquo Toxicon vol 21 no 3 pp 81ndash84 1983

[32] G Faure V T Gowda and R C Maroun ldquoCharacterizationof a human coagulation factor Xa-binding site on Viperidaesnake venom phospholipases A2 by affinity binding studiesand molecular bioinformaticsrdquo BMC Structural Biology vol 7article no 82 2007

[33] J Sribar A Copic A Paris et al ldquoA high affinity acceptor forphospholipase A2 with neurotoxic activity is a calmodulinrdquoTheJournal of Biological Chemistry vol 276 no 16 pp 12493ndash124962001

[34] J Sribar N E Sherman P Prijatelj et al ldquoThe neurotoxic phos-pholipaseA2 associates through a non-phosphorylated bindingmotif with 14-3-3 protein 120574 and 120576 isoformsrdquo Biochemical andBiophysical Research Communications vol 302 no 4 pp 691ndash696 2003

[35] N Vardjan N E Sherman J Pungercar J W Fox F Gubensekand I Krizaj ldquoHigh-molecular-mass receptors for ammody-toxin in pig are tissue-specific isoforms of M-type phos-pholipase A2 receptorrdquo Biochemical and Biophysical ResearchCommunications vol 289 no 1 pp 143ndash149 2001

[36] K Kondo H Toda K Narita and C-Y Lee ldquoAmino acidsequences of three 120573-bungarotoxins (1205733- 1205734- and 1205735-bunga-rotoxins) from Bungarus multicinctus Venom amino acidsubstitutions in the A chainsrdquo The Journal of Biochemistry vol91 no 5 pp 1531ndash1548 1982

[37] M J Sutcliffe CM Dobson and R E Oswald ldquoSolution struc-ture of neuronal bungarotoxin determined by two-dimensionalNMR spectroscopy calculation of tertiary structure usingsystematic homologous model building dynamical simulatedannealing and restrained molecular dynamicsrdquo Biochemistryvol 31 no 11 pp 2962ndash2970 1992

[38] J Halpert and D Eaker ldquoAmino acid sequence of a presynapticneurotoxin from the venom of Notechis scutatus scutatus(Australian tiger snake)rdquo The Journal of Biological Chemistryvol 250 no 17 pp 6990ndash6997 1975

[39] B Westerlund P Nordlund U Uhlin D Eaker and H EklundldquoThe three-dimensional structure of notexin a presynapticneurotoxic phospholipase A2 at 20 A resolutionrdquo FEBS Lettersvol 301 no 2 pp 159ndash164 1992

[40] I I Kaiser J M Gutierrez D Plummer S D Aird and G VOdell ldquoThe amino acid sequence of a myotoxic phospholipasefrom the venomofBothrops asperrdquoArchives of Biochemistry andBiophysics vol 278 no 2 pp 319ndash325 1990

[41] A M Soares V M Rodrigues M I Homsi-Brandeburgo et alldquoA rapid procedure for the isolation of the LYS-49 myotoxin IIfrom bothrops moojeni (caissaca) venom biochemical charac-terization crystallization myotoxic and edematogenic activityrdquoToxicon vol 36 no 3 pp 503ndash514 1998

[42] J R Almeida M Lancellotti A M Soares et al ldquoCoaTx-II anew dimeric Lys49 phospholipase A2 from Crotalus oreganusabyssus snake venom with bactericidal potential Insights intoits structure and biological rolesrdquo Toxicon vol 120 pp 147ndash1582016

[43] V L Bonfim L A Ponce-Soto J C Novello and S MarangonildquoStructural and functional properties of Cr 5 a new Lys49 phos-pholipase A2 homologue isolated from the venom of the snakeCalloselasma rhodostomardquoThe Protein Journal vol 25 no 7-8pp 492ndash502 2006

[44] L A Ponce-Soto B Lomonte J M Gutierrez L Rodrigues-Simioni J C Novello and S Marangoni ldquoStructural andfunctional properties of BaTX a new Lys49 phospholipase A2homologue isolated from the venom of the snake Bothropsalternatusrdquo Biochimica et Biophysica Acta (BBA) - GeneralSubjects vol 1770 no 4 pp 585ndash593 2007

[45] V L Bonfim L A Ponce-Soto D Martins de Souza et alldquoStructural and functional characterization of myotoxin Cr-IV 1 a phospholipase A2 D49 from the venom of the snakeCalloselasma rhodostomardquo Biologicals vol 36 no 3 pp 168ndash176 2008

[46] I KRIZAJ A L BIEBER A RITONJA and F GUBENsEKldquoThe primary structure of ammodytin L a myotoxic phospholi-pase A2 homologue fromVipera ammodytes venomrdquo EuropeanJournal of Biochemistry vol 202 no 3 pp 1165ndash1168 1991

[47] M Sharma J K Iyer N Shih et al ldquoDaboxin p a majorphospholipase A2 enzyme from the indian daboia russeliirusselii venom targets factor x and factor xa for its anticoagulantactivityrdquo PLoS ONE vol 11 no 4 Article ID e0153770 2016

[48] A K Chakraborty R H Hall and A C Ghose ldquoPurificationand characterization of a potent hemolytic toxin with phos-pholipase A2 activity from the venom of Indian Russellrsquos viperrdquoMolecular andCellular Biochemistry vol 237 no 1-2 pp 95ndash1022002

[49] R T Kerns RMKini S Stefansson andH J Evans ldquoTargetingof venom phospholipasesThe strongly anticoagulant phospho-lipase A2 from Naja nigricollis venom binds to coagulationfactor Xa to inhibit the prothrombinase complexrdquo Archives ofBiochemistry and Biophysics vol 369 no 1 pp 107ndash113 1999

[50] RMKini ldquoStructure-function relationships andmechanismofanticoagulant phospholipase A2 enzymes from snake venomsrdquoToxicon vol 45 no 8 pp 1147ndash1161 2005

[51] F J Joubert ldquoNaja mossambica mossambica venom Purifica-tion some properties and the amino acid sequences of threephospholipases A (CM-I CM-II and CM-III)rdquo BBA - ProteinStructure vol 493 no 1 pp 216ndash227 1977

[52] W W Lin P L Chang C Y Lee and F J Joubert ldquoPhar-macological study on phospholipases A2 isolated from Najamossambica mossambica venomrdquo Proceedings of the NationalScience Council Republic of China Part B Life Sciences vol 11no 2 pp 155ndash163 1987

[53] U K Ranawaka D G Lalloo H J de Silva and J WhiteldquoNeurotoxicity in snakebitemdashthe limits of our knowledgerdquo


PLOSNeglected Tropical Diseases vol 7 no 10 Article ID e23022013

[54] J Pungercar and I Krizaj ldquoUnderstanding the molecularmechanism underlying the presynaptic toxicity of secretedphospholipases A2rdquo Toxicon vol 50 no 7 pp 871ndash892 2007

[55] T Petan I Krizaj M H Gelb and J Pungercar ldquoAmmody-toxins potent presynaptic neurotoxins are also highly efficientphospholipase A2 enzymesrdquo Biochemistry vol 44 no 37 pp12535ndash12545 2005

[56] B Lomonte and J Rangel ldquoSnake venomLys49myotoxins fromphospholipases A2 to non-enzymatic membrane disruptorsrdquoToxicon vol 60 no 4 pp 520ndash530 2012

[57] R Otero J Gutierrez M Beatriz Mesa et al ldquoComplications ofBothrops Porthidium andBothriechis snakebites inColombiaA clinical and epidemiological study of 39 cases attended in auniversity hospitalrdquo Toxicon vol 40 no 8 pp 1107ndash1114 2002

[58] J M Gutierrez and C L Ownby ldquoSkeletal muscle degenerationinduced by venom phospholipases A2 insights into the mecha-nisms of local and systemic myotoxicityrdquo Toxicon vol 42 no 8pp 915ndash931 2003

[59] R Milani Junior M T Jorge F P Ferraz de Campos et alldquoSnake bites by the jararacucu (Bothrops jararacussu) clini-copathological studies of 29 proven cases in Sao Paulo StateBrazilrdquo QJM An International Journal of Medicine vol 90 no5 pp 323ndash334 1997

[60] C Montecucco J M Gutierrez and B Lomonte ldquoCellularpathology induced by snake venom phospholipase A2 myotox-ins and neurotoxins common aspects of their mechanisms ofactionrdquo Cellular and Molecular Life Sciences vol 65 no 18 pp2897ndash2912 2008

[61] B Lomonte and J M Gutierrez ldquoPhospholipases A2 fromviperidae snake venoms how do they induce skeletal muscledamagerdquo Acta Chimica Slovenica vol 58 no 4 pp 647ndash6592011

[62] B Lomonte E Moreno A Tarkowski L A Hanson and MMaccarana ldquoNeutralizing interaction between heparins andmyotoxin II a lysine 49 phospholipase A2 from Bothrops aspersnake venom identification of a heparin-binding and cytolytictoxin region by the use of synthetic peptides and molecularmodelingrdquoThe Journal of Biological Chemistry vol 269 no 47pp 29867ndash29873 1994

[63] C E Nuez Y Angulo and B Lomonte ldquoIdentification ofthe myotoxic site of the Lys49 phospholipase A2 from Agk-istrodon piscivorus piscivorus snake venom Synthetic C-terminal peptides from Lys49 but not from Asp49 myotoxinsexert membrane-damaging activitiesrdquo Toxicon vol 39 no 10pp 1587ndash1594 2001

[64] L Chioato E A Aragao T Lopes Ferreira A Ivo de MedeirosL H Faccioli and R J Ward ldquoMapping of the structuraldeterminants of artificial and biological membrane damagingactivities of a Lys49 phospholipase A2 by scanning alaninemutagenesisrdquo Biochimica et Biophysica Acta (BBA) - Biomem-branes vol 1768 no 5 pp 1247ndash1257 2007

[65] H M Verheij M-C Boffa C Rothen M Bryckaert RVerger and G H de Haas ldquoCorrelation of Enzymatic Activityand Anticoagulant Properties of Phospholipase A2rdquo EuropeanJournal of Biochemistry vol 112 no 1 pp 25ndash32 1980

[66] R M Kini ldquoAnticoagulant proteins from snake venoms Struc-ture function and mechanismrdquo Biochemical Journal vol 397no 3 pp 377ndash387 2006

[67] D Saikia R Thakur and A K Mukherjee ldquoAn acidic phos-pholipase A2 (RVVA-PLA2-I) purified from Daboia russelli

venom exerts its anticoagulant activity by enzymatic hydrolysisof plasma phospholipids and by non-enzymatic inhibitionof factor Xa in a phospholipidsCa2+ independent mannerrdquoToxicon vol 57 no 6 pp 841ndash850 2011

[68] S Stefansson R M Kini and H J Evans ldquoThe basic phos-pholipase A2 from Naja nigricollis venom inhibits the pro-thrombinase complex by a novel nonenzymatic mechanismrdquoBiochemistry vol 29 no 33 pp 7742ndash7746 1990

[69] R P Samy P Gopalakrishnakone and V T Chow ldquoTherapeuticapplication of natural inhibitors against snake venom phospho-lipase A2rdquo Bioinformation vol 8 no 1 pp 48ndash57 2012

[70] S Marcussi C D SantrsquoAna C Z Oliveira et al ldquoSnakevenom phospholipase A2 inhibitors Medicinal chemistry andtherapeutic potentialrdquo Current Topics in Medicinal Chemistryvol 7 no 8 pp 743ndash756 2007

[71] A M Soares F K Ticli S Marcussi et al ldquoMedicinal plantswith inhibitory properties against snake venomsrdquo CurrentMedicinal Chemistry vol 12 no 22 pp 2625ndash2641 2005

[72] M Deepa and T Veerabasappa Gowda ldquoPurification andcharacterization of a glycoprotein inhibitor of toxic phospho-lipase from Withania somniferardquo Archives of Biochemistry andBiophysics vol 408 no 1 pp 42ndash50 2002

[73] L Mishra B B Singh and S Dagenais ldquoScientific basis forthe therapeutic use of Withania somnifera (ashwagandha) areviewrdquo Alternative Medicine Review vol 5 no 4 pp 334ndash3462000

[74] D K Machiah and T V Gowda ldquoPurification of a post-synaptic neurotoxic phospholipase A2 from Naja naja venomand its inhibition by a glycoprotein from Withania somniferardquoBiochimie vol 88 no 6 pp 701ndash710 2006

[75] M H Borges A M Soares V M Rodrigues et al ldquoEffectsof aqueous extract of Casearia sylvestris (Flacourtiaceae) onactions of snake and bee venoms and on activity of phos-pholipases A2rdquo Comparative Biochemistry and Physiology - BBiochemistry and Molecular Biology vol 127 no 1 pp 21ndash302000

[76] M H Borges A M Soares V M Rodrigues et al ldquoNeutraliza-tion of proteases from Bothrops snake venoms by the aqueousextract from Casearia sylvestris (Flacourtiaceae)rdquo Toxicon vol39 no 12 pp 1863ndash1869 2001

[77] W L G Cavalcante T O Campos M Dal Pai-Silva et alldquoNeutralization of snake venom phospholipase A2 toxins byaqueous extract of Casearia sylvestris (Flacourtiaceae) inmouseneuromuscular preparationrdquo Journal of Ethnopharmacologyvol 112 no 3 pp 490ndash497 2007

[78] J A Pereanez A C Patino V Nunez and E Osorio ldquoThebiflavonoid morelloflavone inhibits the enzymatic and biolog-ical activities of a snake venom phospholipase A2rdquo Chemico-Biological Interactions vol 220 pp 94ndash101 2014

[79] C A Cotrim S C B De Oliveira E B S Diz Filho et alldquoQuercetin as an inhibitor of snake venom secretory phospho-lipase A2rdquo Chemico-Biological Interactions vol 189 no 1-2 pp9ndash16 2011

[80] C F Bennett S Mong M A Clarke L I Kruse and ST Crooke ldquoDifferential effects of manoalide on secreted andintracellular phospholipasesrdquo Biochemical Pharmacology vol36 no 5 pp 733ndash740 1987

[81] L J Reynolds B P Morgan G A Hite E D Mihelich and EA Dennis ldquoPhospholipase A2 inhibition and modification bymanoaloguerdquo Journal of the American Chemical Society vol 110no 15 pp 5172ndash5177 1988


[82] S L G Rocha B Lomonte A G C Neves-Ferreira et alldquoFunctional analysis of DM64 an antimyotoxic protein withimmunoglobulin-like structure from Didelphis marsupialisserumrdquo European Journal of Biochemistry vol 269 no 24 pp6052ndash6062 2002

[83] I R Leon A G da Costa Neves-Ferreira S L G da RochaM R de Oliveira Trugilho J Perales and R H Valente ldquoUsingmass spectrometry to explore the neglected glycan moieties ofthe antiophidic proteins DM43 and DM64rdquo Proteomics vol 12no 17 pp 2753ndash2765 2012

[84] S Lizano G Domont and J Perales ldquoNatural phospholipase A2myotoxin inhibitor proteins from snakesmammals and plantsrdquoToxicon vol 42 no 8 pp 963ndash977 2003

[85] S Quiros A Alape-Giron Y Angulo and B Lomonte ldquoIsola-tion characterization and molecular cloning of AnMIP a new120572-type phospholipase A2 myotoxin inhibitor from the plasmaof the snake Atropoides nummifer (Viperidae Crotalinae)rdquoComparative Biochemistry and Physiology - B Biochemistry andMolecular Biology vol 146 no 1 pp 60ndash68 2007

[86] C Z Oliveira N A Santos-Filho D L Menaldo et al ldquoStruc-tural and functional characterization of a 120574-type phospholipaseA 2 inhibitor fromBothrops jararacussu Snake PlasmardquoCurrentTopics in Medicinal Chemistry vol 11 no 20 pp 2509ndash25192011

[87] S Lizano B Lomonte J W Fox and J M Gutierrez ldquoBio-chemical characterization and pharmacological properties of aphospholipase A2 myotoxin inhibitor from the plasma of thesnake Bothrops asperrdquo Biochemical Journal vol 326 no 3 pp853ndash859 1997

[88] N Ohkura H Okuhara S Inoue K Ikeda and K HayashildquoPurification and characterization of three distinct types ofphospholipase A2 inhibitors from the blood plasma of the Chi-nese mamushi Agkistrodon blomhoffii siniticusrdquo BiochemicalJournal vol 325 no 2 pp 527ndash531 1997

[89] K Chen L-P Zhong L-Z Chen X Li X Xu and C-H Huang ldquoInvestigation and purification of snake venomsecretory phospholipase A2 inhibitors from sera of somecommon snake species in Jiangxi provincerdquo PharmaceuticalBiotechnology vol 18 no 3 pp 220ndash223 2011

[90] Z Le X Li P Yuan P Liu and C Huang ldquoOrthogonal opti-mization of prokaryotic expression of a natural snake venomphospholipaseA2 inhibitor fromSinonatrix annularisrdquoToxiconvol 108 pp 264ndash271 2015

[91] M E Garcia Denegri S Marunak J S Todaro L A Ponce-Soto O Acosta and L Leiva ldquoNeutralisation of the pharma-cological activities of Bothrops alternatus venom by anti-PLA2IgGsrdquo Toxicon vol 86 pp 89ndash95 2014

[92] J P Rodriguez M De Marzi S Marunak E L MalchiodiL C Leiva and O Acosta ldquoRabbit IgG antibodies againstphospholipase A2 from Crotalus durissus terrificus neutralizethe lethal activity of the venomrdquo Medicina vol 66 no 6 pp512ndash516 2006

[93] L S Fusco J P Rodrıguez F Torres-Huaco et al ldquoP9a(Cdt-PLA2) from Crotalus durissus terrificus as good immunogento be employed in the production of crotalic anti-PLA2 IgGrdquoToxicology Letters vol 238 no 1 pp 7ndash16 2015

[94] M Karakas and W Koenig ldquoVarespladib methyl an oral phos-pholipase A2 inhibitor for the potential treatment of coronaryartery diseaserdquo IDrugs vol 12 no 9 pp 585ndash592 2009

[95] D De Luca A Minucci J Trias et al ldquoVarespladib inhibitssecretory phospholipase A2 in bronchoalveolar lavage of differ-ent types of neonatal lung injuryrdquo Clinical Pharmacology andTherapeutics vol 52 no 5 pp 729ndash737 2012

[96] R S Rosenson M Elliott Y Stasiv and C Hislop ldquoRan-domized trial of an inhibitor of secretory phospholipase A2on atherogenic lipoprotein subclasses in statin-treated patientswith coronary heart diseaserdquo European Heart Journal vol 32no 8 pp 999ndash1005 2011

[97] M Lewin S Samuel J Merkel and P Bickler ldquoVarespladib(LY315920) appears to be a potent broad-spectrum inhibitorof snake venom phospholipase A2 and a possible pre-referraltreatment for envenomationrdquo Toxins vol 8 no 9 article no248 2016

[98] DMora-Obando J Fernandez CMontecucco J M Gutierrezand B Lomonte ldquoSynergism between basic Asp49 and Lys49phospholipase A2 myotoxins of viperid snake venom in vitroand in vivordquo PLoS ONE vol 9 no 10 Article ID e109846 2014

[99] L D Possani B M Martin A Yatani et al ldquoIsolation andphysiological characterization of taicatoxin a complex toxinwith specific effects on calcium channelsrdquo Toxicon vol 30 no11 pp 1343ndash1364 1992

[100] S Xiong Y Luo L Zhong et al ldquoInvestigation of the inhibitorypotential of phospholipase A2 inhibitor gamma from Sinonatrixannularis to snake envenomationrdquo Toxicon vol 137 pp 83ndash912017

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MEDIATORSINFLAMMATION

of


L-amino acid oxidases disintegrins and a few other com-pounds [1] Most svPLA2s hydrolyze glycerophospholipids atthe sn-2 position of the glycerol backbone freeing lysophos-pholipids and fatty acids svPLA2s share 44ndash99 aminoacid identity in their primarily structure which results tohigh similarity in their tertiary structure [8] Based on theirsize location function substrate specificity and calciumrequirement PLA2s are classified into six families svPLA2belongs to the secretory PLA2 (sPLA2) family (groups IA IIAand IIB) [9ndash11] Cobras and kraits rattlesnakes and Gaboonvipers have svPLA2s in groups IA IIA and IIB respectively[8] There are also group IB enzymes which are mainlyfound in mammalian pancreas that have been reported insome snake venoms such as Oxyuranus scutellatus [12]Pseudonaja textilis [13] and Micrurus frontalis frontalis [14]These compounds are conserved in structure andhave similarmolecular masses (sim10ndash20 kDa) 5ndash7 disulfide bonds andanalogous three-dimensional structures [15] InGroup I thereare approximately 115ndash120 residues 7 disulfide bonds (theunique disulfide linking residues 11 and 77) and G IA has acharacteristic surface loop between residues 63 to 67 calledelapidic loop [11] While G IB has a five amino acids residues(residues 62ndash67) extension termed pancreatic loop some GIB snake venom PLA2 even has an eight-residue propeptidesegment in their mature state [13 16] In contrast GroupII has a C-terminal extension the unique disulfide linkingresidues 50 and 137 GIIA have a 7-residue C-terminal exten-sion and seven conserved disulfide bonds while inGroup IIBthe C-terminal extension is 6 residues and only six disulfidesremained in which a universally conserved 61ndash95 disulfideis lacking [11] Furthermore a new subgroup (Lys49 PLA2homologues) can be created through mutation Replacementof the 49th residue (asparagine) with lysine results in aninactive or weakly toxic PLA2 This lysine residue can alsointeractwith other amino acids in the ldquocalcium-binding looprdquoresulting in the loss of calcium-dependent catalytic activity[17 18] Most svPLA2s exist as monomers but some exist incomplexes which mainly exhibit presynaptic neurotoxicitythrough combination of isoenzymes or other proteins [19]

3 PLA2s Are Extensively Distributed inSnake Venom

Mackessy [20] analyzed crude venom from the main cladesof venomous snakes via SDS-PAGE and found that svPLA2sexisted in almost every family (Figure 1) The highestamounts were found in Elapidae Viperidae and Hydrophi-idae The lowest were found in Colubridae (which is usuallynonvenomous) Through the application of transcriptomicsand proteomics we gained a better understanding of venomcomposition and the pharmacological properties of thevenom components [21] Betzel et al found that PLA2smade up 32ndash598 in Viperidae snake venom [22] HoweverBungarus fasciatus venom was found to consist of up to 71of PLA2s [23] Moreover Gutierrez and Lomonte found thatthemost lethal fractions inMicrurus fulvius (family Elapidae)were two PLA2 molecules which represented 334 of thewhole venom [24] To date more than 464 unique svPLA2shave been recorded in UniProtKB database What has been

presented above indicates that PLA2s are abundant and fataltoxins in most snake venoms

4 svPLA2s Have a Wide Spectrum ofPharmacological Effects

Despite producing lysophospholipids and fatty acid proin-flammatorymediators svPLA2s also present a wide spectrumof pharmacological effects in victims (ie neurotoxicitymyotoxicity anticoagulant effects cytotoxicity cardiotoxic-ity and edema Table 1) The diverse toxic effects are tightlyrelated to the multiple functional sites on the surface ofsvPLA2s and their different binding receptors [25]

41 svPLA2 Neurotoxicity Neurotoxic svPLA2s can blockneuromuscular transmission in vertebrate skeletal musclescausing acute neuromuscular weakness and paralysis result-ing in respiratory depression and death [53] NeurotoxicsPLA2s are mainly found in the Elapidae (kraits elapidsand coral snakes) and Viperidae (vipers and rattlesnakes)Their toxicity varies greatly among species ranging from1 120583gkg (Textilotoxin) to 380120583gkg (HDP-2 from Viperanikolskii) [53] Previous studies indicate that there is nocorrelation between toxicity and PLA2 hydrolysis activitysvPLA2 neurotoxicity affects presynaptic nerve terminalsso these compounds are commonly referred as presynap-tic neurotoxins or 120573-neurotoxins (120573-ntxs) [54] 120573-ntxs aremonomers or noncovalent complexes containing 2ndash5 sub-units with at least one PLA2 subunit To our knowledge all 120573-ntxs hydrolyze phospholipids especially anionic lipids (egphosphatidylserine phosphatidic acid and phosphorylatedphosphatidylinositols) which are abundant in the cytosolicleaflets of organelles and the plasmamembrane of eukaryoticcells [55] svPLA2s also bind to special tissue sites to achievetheir neurotoxicity effects The mechanism of svPLA2 neuro-toxicity is still under investigation

42 svPLA2 Myotoxicity svPLA2s can induce acute necrosisof skeletal muscle (myonecrosis) [56] In the envenomationthismyonecrosis can potentially lead to permanent tissue lossor amputation [57] svPLA2 myotoxins are mainly found invenom from Elapidae including sea snakes and Viperidae[58] Depending on the venom these svPLA2s can elicitlocal or systemic myotoxicity Local myotoxicity is mainlyelicited by viperid venom This damage is limited to theregion where the toxin is injected and is often coupledwith hemorrhaging blistering and edema [57 59] Systemicmyotoxicity is elicited by elapid venom (ie some sea snaketerrestrial elapids) This causes muscle damage and a distinctincrease of creatine kinase (CK) activity in plasma and isassociated with renal failure and myoglobinuria [58] Alongwith sharing a highly conserved structure svPLA2myotoxinsare tightly associatedwith neurotoxins Both achieve a similarcellular lesion through membrane perturbation cytosolicCa2+ homoeostasis imbalance and cell degeneration [60]Furthermore some neurotoxic svPLA2s (eg notexin andcrotoxin) cause acute skeletal muscle necrosis adding tosystemic toxic effects (ie rhabdomyolysis) [60]


M (kD)N a

ter

P po

rphy

riacu

s

A a

ntar

cticu

s

H c

yano

cinct

usL

har

dwick

iiL

sem

ifasc

iata

N n

naj

a

N m

elano

leuca

N n

igric

ollis

M f

ulvi

us

Stan

dard

s

B ir

regu

laris

B d

endr

ophi

la

A n

asut

a

T b

iscut

atus

lam

bda

A p

orto

ricen

sis

L m

adag

asca

riens

is



Serine protease()

CRiSP

Phospholipase A2

ree-nger toxins

2000

974

664

556

366

310

210

142

60

35

(a)

T b

orne

ensis

T p

unice

us

T st

ejneg

eri

T fl

avov

iridi

sT

pur

pure

omac

ulat

us

D ru

sselli

i rus

selli

i

C rh

odos

tom

a

A n

itsch

iiC

resim

us

E ca

rinat

us so

chur

eki

B ga

boni

ca ga

boni

ca

B a

rieta

ns

V ra

ddei

V a

mm

odyt

es

C ti

gris

C tr

ansv

ersu

s

stan

dard

s



Serine proteases

Metalloprotease-PI

PLA2C-type lectin

CRiSP

Disintegrins

Myotoxins()

2000

974

664

556

366

310

210

142

60

35

(b)







ityof

snakev

enom

PLA2s

Nam

eSn

akes

pecies

Structuralfeatures

subtyp

eaTo

xicitie

sLethality

inmou

se(120583gkg)b

Bind

ingproteins

intissuec

PLA2activ

ity(120583molm

inm

gtoxin)

dRe

ference

Neurotoxin

Crotoxin

Crotalus

durissusterrifi

cus

Heterod

imericA

IIA-sPL

A2-like

Neurotoxicitym


60ndash240

(iv)

Crocalbin

CaM

85[26]

BIIA

-sPL

A2

MsPLA2-I

Micr

urus

spixii

Mon

omericIA-

PLA2

Neurotoxicitym

yotoxicityantiplasm

odialactivity

edem

and

nAchR

Yes

[27]

Taipoxin

Oxyuranus

scutellatus

Trim

eric120572

IAtoxic120573

IA-

sPLA2lik

e120574IB-sPLA2

glycosylated

Presyn

aptic

neurotoxicity

2(iv)

M-sPL

A2R

NP

TCBP

-49

04

[28ndash30]

cytotoxicity

Textilo

toxin

Pseudona

jatextilis

Pentam

ericA

BandCareIA-

sPLA2D2


Presyn

aptic

neurotoxicity

1(iv)

M-sPL

A2R

32

[132831]

Ammod

ytoxin

Vipera

ammodytes

Mon

omericIIA-sPL

A2

Presyn

aptic

neurotoxicity

21

(iv)

M-sPL

A2R

CaMP

DI

FXa14-3-3

proteins

280

[32ndash35]

antic

oagu

lant

120573-Bun

garotoxin

Bungarus

multicinctus

Dim

ericA

IA-

sPLA2

Presyn

aptic

neurotoxicity

19ndash130

(ip)

v-d

K+channel

61[3637]

S-Slin

kedto

subu

nitB

BPT

I-lik

e

Notexin

Notechisscutatus

Mon

omericIA-sPL

A2(A

sp49)


neph

rotoxicity

17(iv)

nd

1390

[3839]

Myotoxin

MyotoxinIII

Bothrops

asper

Dim

ericIIA

-sPL

A2(A

sp49)

Myotoxicity

470(iv)

nd

750

[40]

antic

oagu

lantedema

MyotoxinII

Bmoojen

iMon

omericIIA-sPL

A2(Lys49)

Myotoxicityedema

7600

(ip)

nd

Non

e[41]

CoaTx

-II

Crotalus

oreganus

abyssus

Dim

ericIIA-sPL

A2(Lys49)


acteria

lactivity

nd

nd

Non

e[42]

Cr5

Calloselasm

arhodostoma

Mon

omericIIA-sPL

A2(Lys49)

Cytotoxicitym

yotoxicityedema

70(icv

)nd

Non

e[43]

BaTX

Bothrops

alternatus

Mon

omericIIA

-sPL

A2(Lys49)

Cytotoxicitym


7000

(iv)

nd

Non

e[44]

Cr-IV1

Calloselasm

arhodostoma

Mon

omericIIA-sPL

A2(A

sp49)


edema

70(icv

)nd

0014

[45]

Ammod

ytin

LVipera

ammodytes

Mon

omericIIA-sPL

A2(Ser49)

Myotoxicity

3600

(ip)

nd

Non

e[46]

Anticoagulant

Daboxin

PDaboiarusse

liiMon

omericIA-

sPLA2

Strong

antic

oagu

lant

nd

FXFXa

1140

[47]

RVV-PFIIc1015840

Drusselii

Mon

omericIIA-sPL

A2(A

sp49)

Anticoagu

lant

100(ip)

nd

Yes

[48]

CM-IV

Najanigrico

llis

Mon

omericIIA-sPL

A2(A

sp49)

Strong

lyantic

oagu


180(ip)

FXaFV

IIaYes

[4950]

CM-II

Najamossambica

Mon

omericIA-

sPLA2

Weakanticoagu

lantm

yotoxicityn

eurotoxicity

nd

TFFVII

Yes

[5152]

a BPT

Ibo

vine

pancreatictrypsin

inhibitor

b icv

intracerebroventric

ularivintraveno

usicintraciste

rnalipintraperito

nealn

dno

tdeterminedcCa

Mcalmod

ulinN

Pneuron

alpentraxin

PDIprotein

disulfide

isomeraseTC


protein49M

-sPL

A2R

M-ty


coagulationfactor

XaFX

bloo

dcoagulationfactor

XTF

tissue

factorFVIIblood

coagulationfactor

VIIFVIIa

blood

coagulationfactor

VIIa

v-d


K+channels

d pho

spho

lipaseA2activ

ityisin120583molm

inm

gof

toxin

Yesoriginalresearch

paperd

oesn

otshow

phosph

olipaseA2activ

ityin

concretenu

mbero

rnot

in120583molm

inm

gof

toxin

Non

eallP

LA2ho

mologuesa

rehere

considered

tobe


eAd

aptedfro

m[5051]
























6 Conclusions









Acknowledgments


References














































































































Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of


M (kD)N a

ter

P po

rphy

riacu

s

A a

ntar

cticu

s

H c

yano

cinct

usL

har

dwick

iiL

sem

ifasc

iata

N n

naj

a

N m

elano

leuca

N n

igric

ollis

M f

ulvi

us

Stan

dard

s

B ir

regu

laris

B d

endr

ophi

la

A n

asut

a

T b

iscut

atus

lam

bda

A p

orto

ricen

sis

L m

adag

asca

riens

is



Serine protease()

CRiSP

Phospholipase A2

ree-nger toxins

2000

974

664

556

366

310

210

142

60

35

(a)

T b

orne

ensis

T p

unice

us

T st

ejneg

eri

T fl

avov

iridi

sT

pur

pure

omac

ulat

us

D ru

sselli

i rus

selli

i

C rh

odos

tom

a

A n

itsch

iiC

resim

us

E ca

rinat

us so

chur

eki

B ga

boni

ca ga

boni

ca

B a

rieta

ns

V ra

ddei

V a

mm

odyt

es

C ti

gris

C tr

ansv

ersu

s

stan

dard

s



Serine proteases

Metalloprotease-PI

PLA2C-type lectin

CRiSP

Disintegrins

Myotoxins()

2000

974

664

556

366

310

210

142

60

35

(b)







ityof

snakev

enom

PLA2s

Nam

eSn

akes

pecies

Structuralfeatures

subtyp

eaTo

xicitie

sLethality

inmou

se(120583gkg)b

Bind

ingproteins

intissuec

PLA2activ

ity(120583molm

inm

gtoxin)

dRe

ference

Neurotoxin

Crotoxin

Crotalus

durissusterrifi

cus

Heterod

imericA

IIA-sPL

A2-like

Neurotoxicitym


60ndash240

(iv)

Crocalbin

CaM

85[26]

BIIA

-sPL

A2

MsPLA2-I

Micr

urus

spixii

Mon

omericIA-

PLA2

Neurotoxicitym

yotoxicityantiplasm

odialactivity

edem

and

nAchR

Yes

[27]

Taipoxin

Oxyuranus

scutellatus

Trim

eric120572

IAtoxic120573

IA-

sPLA2lik

e120574IB-sPLA2

glycosylated

Presyn

aptic

neurotoxicity

2(iv)

M-sPL

A2R

NP

TCBP

-49

04

[28ndash30]

cytotoxicity

Textilo

toxin

Pseudona

jatextilis

Pentam

ericA

BandCareIA-

sPLA2D2


Presyn

aptic

neurotoxicity

1(iv)

M-sPL

A2R

32

[132831]

Ammod

ytoxin

Vipera

ammodytes

Mon

omericIIA-sPL

A2

Presyn

aptic

neurotoxicity

21

(iv)

M-sPL

A2R

CaMP

DI

FXa14-3-3

proteins

280

[32ndash35]

antic

oagu

lant

120573-Bun

garotoxin

Bungarus

multicinctus

Dim

ericA

IA-

sPLA2

Presyn

aptic

neurotoxicity

19ndash130

(ip)

v-d

K+channel

61[3637]

S-Slin

kedto

subu

nitB

BPT

I-lik

e

Notexin

Notechisscutatus

Mon

omericIA-sPL

A2(A

sp49)


neph

rotoxicity

17(iv)

nd

1390

[3839]

Myotoxin

MyotoxinIII

Bothrops

asper

Dim

ericIIA

-sPL

A2(A

sp49)

Myotoxicity

470(iv)

nd

750

[40]

antic

oagu

lantedema

MyotoxinII

Bmoojen

iMon

omericIIA-sPL

A2(Lys49)

Myotoxicityedema

7600

(ip)

nd

Non

e[41]

CoaTx

-II

Crotalus

oreganus

abyssus

Dim

ericIIA-sPL

A2(Lys49)


acteria

lactivity

nd

nd

Non

e[42]

Cr5

Calloselasm

arhodostoma

Mon

omericIIA-sPL

A2(Lys49)

Cytotoxicitym

yotoxicityedema

70(icv

)nd

Non

e[43]

BaTX

Bothrops

alternatus

Mon

omericIIA

-sPL

A2(Lys49)

Cytotoxicitym


7000

(iv)

nd

Non

e[44]

Cr-IV1

Calloselasm

arhodostoma

Mon

omericIIA-sPL

A2(A

sp49)


edema

70(icv

)nd

0014

[45]

Ammod

ytin

LVipera

ammodytes

Mon

omericIIA-sPL

A2(Ser49)

Myotoxicity

3600

(ip)

nd

Non

e[46]

Anticoagulant

Daboxin

PDaboiarusse

liiMon

omericIA-

sPLA2

Strong

antic

oagu

lant

nd

FXFXa

1140

[47]

RVV-PFIIc1015840

Drusselii

Mon

omericIIA-sPL

A2(A

sp49)

Anticoagu

lant

100(ip)

nd

Yes

[48]

CM-IV

Najanigrico

llis

Mon

omericIIA-sPL

A2(A

sp49)

Strong

lyantic

oagu


180(ip)

FXaFV

IIaYes

[4950]

CM-II

Najamossambica

Mon

omericIA-

sPLA2

Weakanticoagu

lantm

yotoxicityn

eurotoxicity

nd

TFFVII

Yes

[5152]

a BPT

Ibo

vine

pancreatictrypsin

inhibitor

b icv

intracerebroventric

ularivintraveno

usicintraciste

rnalipintraperito

nealn

dno

tdeterminedcCa

Mcalmod

ulinN

Pneuron

alpentraxin

PDIprotein

disulfide

isomeraseTC


protein49M

-sPL

A2R

M-ty


coagulationfactor

XaFX

bloo

dcoagulationfactor

XTF

tissue

factorFVIIblood

coagulationfactor

VIIFVIIa

blood

coagulationfactor

VIIa

v-d


K+channels

d pho

spho

lipaseA2activ

ityisin120583molm

inm

gof

toxin

Yesoriginalresearch

paperd

oesn

otshow

phosph

olipaseA2activ

ityin

concretenu

mbero

rnot

in120583molm

inm

gof

toxin

Non

eallP

LA2ho

mologuesa

rehere

considered

tobe


eAd

aptedfro

m[5051]
























6 Conclusions









Acknowledgments


References














































































































Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of




ityof

snakev

enom

PLA2s

Nam

eSn

akes

pecies

Structuralfeatures

subtyp

eaTo

xicitie

sLethality

inmou

se(120583gkg)b

Bind

ingproteins

intissuec

PLA2activ

ity(120583molm

inm

gtoxin)

dRe

ference

Neurotoxin

Crotoxin

Crotalus

durissusterrifi

cus

Heterod

imericA

IIA-sPL

A2-like

Neurotoxicitym


60ndash240

(iv)

Crocalbin

CaM

85[26]

BIIA

-sPL

A2

MsPLA2-I

Micr

urus

spixii

Mon

omericIA-

PLA2

Neurotoxicitym

yotoxicityantiplasm

odialactivity

edem

and

nAchR

Yes

[27]

Taipoxin

Oxyuranus

scutellatus

Trim

eric120572

IAtoxic120573

IA-

sPLA2lik

e120574IB-sPLA2

glycosylated

Presyn

aptic

neurotoxicity

2(iv)

M-sPL

A2R

NP

TCBP

-49

04

[28ndash30]

cytotoxicity

Textilo

toxin

Pseudona

jatextilis

Pentam

ericA

BandCareIA-

sPLA2D2


Presyn

aptic

neurotoxicity

1(iv)

M-sPL

A2R

32

[132831]

Ammod

ytoxin

Vipera

ammodytes

Mon

omericIIA-sPL

A2

Presyn

aptic

neurotoxicity

21

(iv)

M-sPL

A2R

CaMP

DI

FXa14-3-3

proteins

280

[32ndash35]

antic

oagu

lant

120573-Bun

garotoxin

Bungarus

multicinctus

Dim

ericA

IA-

sPLA2

Presyn

aptic

neurotoxicity

19ndash130

(ip)

v-d

K+channel

61[3637]

S-Slin

kedto

subu

nitB

BPT

I-lik

e

Notexin

Notechisscutatus

Mon

omericIA-sPL

A2(A

sp49)


neph

rotoxicity

17(iv)

nd

1390

[3839]

Myotoxin

MyotoxinIII

Bothrops

asper

Dim

ericIIA

-sPL

A2(A

sp49)

Myotoxicity

470(iv)

nd

750

[40]

antic

oagu

lantedema

MyotoxinII

Bmoojen

iMon

omericIIA-sPL

A2(Lys49)

Myotoxicityedema

7600

(ip)

nd

Non

e[41]

CoaTx

-II

Crotalus

oreganus

abyssus

Dim

ericIIA-sPL

A2(Lys49)


acteria

lactivity

nd

nd

Non

e[42]

Cr5

Calloselasm

arhodostoma

Mon

omericIIA-sPL

A2(Lys49)

Cytotoxicitym

yotoxicityedema

70(icv

)nd

Non

e[43]

BaTX

Bothrops

alternatus

Mon

omericIIA

-sPL

A2(Lys49)

Cytotoxicitym


7000

(iv)

nd

Non

e[44]

Cr-IV1

Calloselasm

arhodostoma

Mon

omericIIA-sPL

A2(A

sp49)


edema

70(icv

)nd

0014

[45]

Ammod

ytin

LVipera

ammodytes

Mon

omericIIA-sPL

A2(Ser49)

Myotoxicity

3600

(ip)

nd

Non

e[46]

Anticoagulant

Daboxin

PDaboiarusse

liiMon

omericIA-

sPLA2

Strong

antic

oagu

lant

nd

FXFXa

1140

[47]

RVV-PFIIc1015840

Drusselii

Mon

omericIIA-sPL

A2(A

sp49)

Anticoagu

lant

100(ip)

nd

Yes

[48]

CM-IV

Najanigrico

llis

Mon

omericIIA-sPL

A2(A

sp49)

Strong

lyantic

oagu


180(ip)

FXaFV

IIaYes

[4950]

CM-II

Najamossambica

Mon

omericIA-

sPLA2

Weakanticoagu

lantm

yotoxicityn

eurotoxicity

nd

TFFVII

Yes

[5152]

a BPT

Ibo

vine

pancreatictrypsin

inhibitor

b icv

intracerebroventric

ularivintraveno

usicintraciste

rnalipintraperito

nealn

dno

tdeterminedcCa

Mcalmod

ulinN

Pneuron

alpentraxin

PDIprotein

disulfide

isomeraseTC


protein49M

-sPL

A2R

M-ty


coagulationfactor

XaFX

bloo

dcoagulationfactor

XTF

tissue

factorFVIIblood

coagulationfactor

VIIFVIIa

blood

coagulationfactor

VIIa

v-d


K+channels

d pho

spho

lipaseA2activ

ityisin120583molm

inm

gof

toxin

Yesoriginalresearch

paperd

oesn

otshow

phosph

olipaseA2activ

ityin

concretenu

mbero

rnot

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6 Conclusions









Acknowledgments


References














































































































Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of
























6 Conclusions









Acknowledgments


References














































































































Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of







Acknowledgments


References














































































































Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of





















































































Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of

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