molecular recognition at the lipid-water interface -...

Burkhard KönigDepartment of Chemistry and PharmacyDr. Max MustermannReferat Kommunikation & Marketing Verwaltung

Burkhard König

Molecular recognition at the lipid-water interface

Burkhard KönigDepartment of Chemistry and Pharmacy

The lipid-water interface is a very special place . . .

HO

H HO

H

HO

H

HO

H HO

H

HO

H

HO

H

HO

H

HO

H

HO

H

HO H

HO

H

HO H

HO H

HO

HH

OH

HO

H

• 3-D interactionsreduced to 2-D

• less degrees offreedom

• changes in entropy

• intramolecular forcesand physical constantslike pKa change dramatically


. . . for molecular recognition

HO

H HO

H

HO

H

HO

H HO

H

HO

H

HO

H

HO

H

HO

H

HO

H

HO H

HO

H

HO H

HO H

HO

HH

OH

HO

H


Chemical Bonds1000 100 10 1[KJ/mol]

Van-der-Waals0.5 - 5

N Cl

C N Cl

-N...Cl-3 - 5 (calcd.)

C C

840

C N

C O

C CC C

C Si

C F

I IO H

C O

H F

890

610

740415

486360

465151

301565

AlH3160

covalent, metallic, ionic (1000 - 50) H OH O

H

H21

CT complexes5 - 50

inclusion inhydrophobic cavity 30

hydrogen bonds8 - 40

dipole-dipole 4 - 25

BO

ON

NN

NCu

+

F3CSO3-

NNZn

2+

NN

OH2

2 ClO4-reversible

coordinative bonds

reversible covalentbonds


Metal-ion tightly coordinated in a multidentate ligand

Lewis-basic guest molecule

+

Reversible coordinative bonds

Use of reversible coordinative bonds in molecular recognition: M. Kruppa, BK, Chem. Rev. 2006, 106, 3520

HSAB-guided selectivity


Reversible coordinative bonds

Me

O

ON

O

O

O

O

H2O

H2O

O

OR

NCu

O

H2OO

OH2

H2O O

ONHR

( )6

Copper imidodiacetato (IDA) complexbinds histidine andpyrophosphate

Nitrilo triacetato (NTA) metal complexbinds histidine

N

N

NHH

HHZn

2+ N

OH2 2 ClO4-

Zinc-cyclen complexbinds phosphates and imides


Dipeptide Chemosensors

NH3

NH

O

O

NH2O

N

NH

O

O

OO

OOO

OOO

N

N

Cu

O

OH2

O

O

O

O

O

O

O OO

O

OOO

N

NCu

O

OH2

O

OH2

OH2

O

O

+

His-Lys-OMeHEPES, pH 7.5

Receptor Receptor + His-Lys-OMe

log K = 4.22 ± 0.05

M. Kruppa, C. Mandl, S. Miltschitzky, BK J. Am. Chem. Soc.2005, 127, 3362;

A. Riechers, S. Stadlbauer, A. Späth, BK, Chem. Eur. J. 2008, 14, 2536;A. Grauer, A. Riechers, S. Ritter, BK, Chem. Eur. J. 2008, 14, 8922.


Nucleotide sensing

N

NN

NH H

H

Zn2+

2 Cl-NH

O

F. Schmidt, S. Stadlbauer, BK Dalton Trans., 2010, in print.

[ZnCycPyr] = 0.33 mM;

[analyt] = 0.17 mM

50 mMHEPES, 154 mM NaCl, pH

7.4exc @ 360

nm


SDS PAGE Phosphoprotein staining

A. Riechers, F. Schmidt, S. Stadlbauer, BK, Bioconj. Chem. 2009, 20, 804–807; patent pending.

A

B

Co

om

assi

e

α-casein dephosphorylated

500 ng 15 ng


SDS PAGE DNA staining

AT

AG

C

F. Schmidt, J. Schmidt, A. Riechers, S. Haase, A.-K. Bosserhoff, J. Heilmann, BK Nucleos. Nucleot. Nucl., 2010, in print.


Photochromic enzyme inhibitorlight 1

light 2

SS SS

312 nm

> 434 nm


Photochromic enzyme inhibitor

Cu

O

ON

O

O

O

H2O

H2O

SSN

O O

H

SO

ONH2

Cu

O

ON

O

O

O

H2O

H2O

SSN

O O

H

SO

ONH2

312 nm> 434 nm

H2

H2

D. Vomasta, C. Högner, N. R. Branda, BK, Angew. Chem. Int. Ed. 2008, 47, 7644;D. Vomasta, A. Innocenti, BK, C. T. Supuran, Bioorg. Med. Chem. Lett. 2009, 19, 1283.


Cu

O

ON

O

O

O

H2O

H2O

SSN

O O

H

SO

ONH2

Cu

O

ON

O

O

O

H2O

H2O

SSN

O O

H

SO

ONH2

NH2

SO

ONH2

H2

H2

Human Carbonic Anhydrase I

CO2 + H2O H2CO3

D. Vomasta, C. Högner, N. R. Branda, BK, Angew. Chem. Int. Ed. 2008, 47, 7644.


Photochromic enzyme inhibitor


Gold surface covered by alkane thiols

Amphiphilic receptors transferred as Langmuir Blodget monolayer

M. Kalinina, BK, M. Subat, S.L. Selector et al. Langmuir 2007, 23, 2517.


M. Kalinina, BK, M. Subat et al. Angew. Chem.2006, 118, 5466.

Stepwise adsorption of nucleotidesSurface plasmon resonance (SPR) sensograms


M. Kalinina, BK, M. Subat et al. Angew. Chem. 2006, 118, 5466; D. S. Turygin, BK, M. A. Kalinina et al., Chem. Eur. J. 2010, in print.

Stepwise adsorption of nucleotidesHydrogen bond guided self assembly at the surface


Functionalized double layers


roll up !

Functionalized double layers yield . . .


. . surface modified vesicles

Towards artificial antibodies• Nanometer size• Multipoint interaction


Surface modified polydiacetylene vesicles

D. Amilan Jose, Stefan Stadlbauer, BK, Chem. Eur. J. 2009, 15, 7404.



Sonication @ 70°C Syringe filter

Store @ 0°C Dispersion Blue coloured vesicles;

180-250 nm

Irradiation254nm



D. Amilan Jose, Stefan Stadlbauer, BK, Chem. Eur. J. 2009, 15, 7404.


Polydiacetylene vesicles on filter paper


Color change by aggregation

added ppi added H2PO4-

HN

O

NH

8

9

N

N

N

NZn

H H

HN

N

N

N

N

N

NZn

H H

H

4+


Polydiacetylen vesicles responding to metal ions


Polydiacetylen vesicles responding to metal ions

LS-Terpy

LS-DPA

LS-DP

LS-DEA

Li+ Na+ K+ Mn2+ Cu2+ Ni2+ Co2+ Cr2+ Zn2+ Cd2+ Ag+ Hg2+

D. A. Jose, BK, Org. Biomol. Chem. 2010, 655-662


Surface modified small unilamellar vesicles. . .


. . . allow indicator displacement assays

emis

sion

en

hanc

emen

t [%

]

coumarin methylSulfonate as indicator

B. Gruber, S. Stadlbauer, K. Woinaroschy, BK, Org. Biomol. Chem. 2010, 8, 3704.


Embedded luminescent amphiphilic receptors . . .

No emission response to analytesin homogeneous solution B. Gruber, S. Stadlbauer, K. Woinaroschy,

BK, Org. Biomol. Chem. 2010, 8, 3704.


. . . respond to the presence of ATP and PPi . . . .


N

NN

N

NH2

O

OH

OPOO-

OPOPO

O-

O

O-

-O

pH 7, Hepes, lg K = 6.60 – 5 eq. of pyrophosphate

emis

sion

inte

nsity


. . . and detect phosphorylated proteins



Co-embedding of binding site and dye . . .

B. Gruber, S. Stadlbauer, A. Späth, S. Weiss, M. Kalinina, BK, Angew. Chem. 2010, DOI: 10.1002/anie.201001101.


…. facilitates quick optimization of properties



Coumarin Fluorescine

response to PPi


receptor(s) analyte lg K (± 0.1)

1 PPi 6.2

1 pSer 5.9

2 4-Me-Im 7.1

2 His-OMe 7.1

1 + 2 PPi 6.0

1 + 2 pSer 5.8

1 + 2 4-Me-Im 6.2

1 + 2 His-OMe 6.1

1 + 2 + 3 PPi 6.1

1 + 2 + 3 4-Me-Im 6.0

1 + 2 + 3 Gly-OMe 5.8

Additive receptor response to analytes


1 = affinity for phosphate anions

2 = affinity for imidazole

3 = affinity for ammonium ions


Lipid – water interfaces with embeddedsynthetic receptors and dyes allow . . .

guided self-assembly at surfaces

facile preparation of analytical nanoparticles

and of liposomes with catalytic activity

Our vision: Preparation of highly selective nanoparticlesfür protein surface recognition Synthetic antibodiesand of Nanocatalystsfor selective transformation of biomolecules

Thank you !The group 2010:

Stefan Füldner,, Carolin Fischer, Stefan Weiß, Carolin Russ,,Andreas Hohenleutner, Benjamin Gruber, Susanna Schmidtbauer, Peter Raster, Josef Herrmann, Cristian Puentes, Mouchumi Bhuyan, Michael Dobmeier, Karin Lehner, Dr. Evgeny Katayev, Dr. Tatiana Mitkina, Dr. Anna Berlicka, Christoph Harlander, Peter Schroll, Veronika Flügel, Stefan Balk, Tobias Lang, Natasha Kuzmanovic, Matthias Wrobel, Andreas Müller, Durag Prasa, Sandip Bhowmik

Alumni club:Dr. Harald Zieg, Dr. Mario Pelka, Dr. Martin Rödel, Mirjam Sax,Dr. Clemens Horn, Dr. Stefanie Leue,Thorsten Graf, Bernhard Lerche,Daniela Fischer, Andreas Fuchs,Dr. Tom Fricke, Dr. Wolfgang Pitsch,Natascha Naarmann, Dr. Hans-Christoph Gallmeier, Dr. Roland Reichenbach-KlinkeDr. Mike Kercher, Miriam Kemter, Dr. Radek Cibulka, Dr. Katerina Cernovska, Dr. Valery Kozhevnikov, Dr. Maria Hechavarria Fonseca, Dr. Michael Klein, Martin Eiblmeier, Dr. Christoph Bonauer; Dr. Christian Mandl; Björn Bartel; Dr. Thomas Walenzyk; Dr. Stefan Miltschitzky; Dr. Thomas Suhs; Dr. Michael Kruppa; Dr. Veronika Michlova; Dr. Xiaoqiang Li, Dr. Xuqin Li; Dr. Georg Dirschl; Dr. Jiri Svoboda; Dr. Kristina Woinaroschy; Dr. Stefan Ritter; Dr. Giovanni Imperato; Barbara FreundDaniel Gölderniz, Dr. Maity Prantik, Dr. Daniel Vomasta, Dr. Andreas Grauer, Dr. Michael Egger, Dr. Jens Geduhn, Dr. Harald Schmaderer, Dr. Florian Ilgen, Dr. Stefan Stadlbauer, Dr. AmilanJose, Dr. Muruganantham Rajendran (Anand), Ina Ehlers, Markus Daerr, Florian Kinzl, Robert Lechner, Andreas Späth, Alexander Riechers

Collaborations:Prof. Maria Kalinina, MoskauProf. Hans R. Kalbitzer, RegensburgProfs Paul Hanson, Jeff Aubé, KansasProf. Uday Maitra, Bangalore

Support:Universität Regensburg, Fonds derChemischen Industrie, DFG, VolkswagenStiftung, Deutsche BundesstiftungUmwelt, DAAD, Alexander von HumboldtStiftung, Evonik Stiftung, FachagenturNawaros, bayrische Forschungsstiftung

The City of Regensburg

http://www.oc-praktikum.de

molecular recognition at the lipid-water interface -...

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