Biomembranes: Structure and Biophysical PropertiesCHM 6304 section 5731Spring 2006

meeting time MWF 8th (3-3:50)would like to move meeting to CLB 313 (outside my office)

Grading:

20% Class Participation30% In-class Presentations25% Midterm25% Final

No Classes:

Jan 16thFeb 20-24thMarch 13-17April 12th

Student Presentations (10 registered folks, sign up for 2 - 20 minutetalks)

•Presentations focus on a topic and a technique.•Presentations will be on Fridays•I as well as the other students will grade you•Presentations are based upon literature. You MUST send out a pdf file one week prior to presentation - loss of 20% grade if not.•Part of in-class grade will come from questions and discussions generated from each presentation.•See syllabus for list of journals. (not inclusive, just a guide)

Week # Date Topic/Technique Presenter 1 Presenter 2 3 Jan 27 Lipid mesophases

DSC/NMR /X-ray

4 Feb 3 Detergent-lipid phases Dynamic light scattering

5 Feb 10 Anesthetics-small molecule partitioning EPR /NMR

6 Feb 17 Cholesterol/lipid rafts Fluorescence :FRET/FRAP etc

8 Mar 3 Membrane Enzymes/Lipid Assymmetry Assays/chemical modification

11 Mar 24 Antibiotic peptides/cell penetrating peptides Isothermal titration calorimetry

12 Mar 31 Membrane proteins- channels/pores Structure-reconstitution

13 Apr 7 Membrane proteins receptors/signal transduction

14 Apr 14 Membrane fusion/membrane curvature EPR/fluorescence

15 Apr 21 Lipid phases for crystallization/ x-ray

Hopefully lecture will always be ahead of the topics for presentations. Changing of topics is open for discussion, but must first be approved by me.

There is no required text book.

Recommended Texts:Biomembranes: Molecular Structure and Function

Robert B. Gennis

The Structure of Biological MembranesPhilip L. Yeagle

Additional Recommended Texts:The Membranes of Cells

Philip L. Yeagle

The Phospholipid Manual

PLEASE DO NOT RECALL BOOKS FROM LIBRARYI will provide scanned pdf files,material will be supplemented with recent lit and reviews (pdf)

To get pdf files:

you MUST send me your email address so I can make a list.I will send out email with the link.

I will set up a website/ftp site for the course. All materials will be deposited with folders that are dated.

Lecture notes will be handed out prior to class.They will NOT be posted on-line.

Lipid Structure and Classification

amphipathic

Polar headgroup

Non-polar chains

Biomembranes are comprised predominantly of lipids

Non-saponifiableContain fatty acids Do not Contain fatty acids

Saponifiable

sterols

Lipid Classification/Structure Continued

OH

Fatty acids can be ester or ether linked

Phosphoglycerides

Sphingosine

Ceramide

Phosphatydic acid

Note different charges of head groups-

learn abbreviations

General Structure of Glycerophospholipids

Fatty acid diversity

saturatedcis

unsaturated transunsaturated

Oleic acid

9,12-linoleic acid

-linoleic acid

-linoleic acid

arachidonic acid

Polyunsaturated fatty acids: PUFAs

cardiolipin

diacylglycerol

dialkylglycerol

triacylglycerol

trialkylglycerol

Cholesteryl oleate

Diphosphatidylglycerol DPG

Misc. fats

(LDL)

Sphingolipids

Sphingosine

N-Acyl-sphingosine or Ceremide

Sphingomyelin

1-O--D-galactosyl-sphingosine orpsychosine

Glycolipids

Common in plants, rare in animals

-D-Galactosyl-diacylglycerol

glycoglycerolipids glycosphingolipids

cerebrosides

gangliosides

sialic acid

Lipopolysaccharides

Lipid ALipid X

endotoxin

Sterols

13

149810

171211

15

16

756

20CH318

CH3191

2322

4

CH321

24

2

3H

H

H

H25

CH327

CH326

cholestane

TLC Analysis of Lipids

How to make this quantitative? Phosphate Assays...

• Separation is based upon polarity• Visualization:

• Sulfuric acid/charring• Iodine vapor• Fluorescence

Quantify Lipids: Phosphate assays.Scrape spots from plates, extract

lipids with solvent,Decompose lipids into inorganic

phosphate, detect with coloring agent (based upon molybdate)

Read blue color at 660nm against standards.

Colors for specific head group identification:

PC in phosphoglycerides and sphingomylin- Dragendorgg’s reagent (KI, Bi)dark blue spots = phosphateneutral or glyco do not stain

Amino Groups (PE and PS)ninhydrin spray, positive is pink-purple

Glycolipids-naphthol give purple spots

Phospholipase Digestions Followed by GC Analysis