Transcript lecture 1 - Fanucci Group

Biomembranes: Structure and Biophysical Properties
CHM 6304 section 5731
Spring 2006
meeting time MWF 8th (3-3:50)
would like to move meeting to CLB 313 (outside my office)
Grading:
No Classes:
20% Class Participation
30% In-class Presentations
25% Midterm
25% Final
Jan 16th
Feb 20-24th
March 13-17
April 12th
Student Presentations (10 registered folks, sign up for 2 - 20 minute
talks)
•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
3
Jan 27
4
Feb 3
5
Feb 10
6
Feb 17
8
Mar 3
11
Mar 24
12
Mar 31
13
Apr 7
14
Apr 14
15
Apr 21
Lipid mesophases
DSC/NMR /X-ray
Detergent-lipid phases
Dynamic light scattering
Anesthetics-small molecule
partitioning
EPR /NMR
Cholesterol/lipid rafts
Fluorescence :FRET/FRAP
etc
Membrane Enzymes/Lipid
Assymmetry
Assays/chemical
modification
Antibiotic peptides/cell
penetrating peptides
Isothermal titration
calorimetry
Membrane proteinschannels/pores
Structure-reconstitution
Membrane proteins
receptors/signal transduction
Membrane fusion/membrane
curvature
EPR/fluorescence
Lipid phases for
crystallization/ x-ray
Presenter 1
Presenter 2
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 Membranes
Philip L. Yeagle
Additional Recommended Texts:
The Membranes of Cells
Philip L. Yeagle
The Phospholipid Manual
PLEASE DO NOT RECALL BOOKS FROM LIBRARY
I 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.
Biomembranes are comprised predominantly of lipids
Lipid Structure and Classification
amphipathic
sterols
Polar headgroup
Non-polar chains
Saponifiable
Contain fatty acids
Non-saponifiable
Do not Contain fatty acids
Lipid Classification/Structure Continued
Phosphoglycerides
Fatty acids can be ester or ether linked
Phosphatydic acid
Sphingosine
OH
Ceramide
General Structure of Glycerophospholipids
Note different charges of head groupslearn abbreviations
Fatty acid diversity
Oleic acid
9,12-linoleic acid
a-linoleic acid
g-linoleic acid
saturated
cis
unsaturated
trans
unsaturated
arachidonic acid
Polyunsaturated fatty acids: PUFAs
Misc. fats
cardiolipin
Diphosphatidylglycerol DPG
diacylglycerol
dialkylglycerol
triacylglycerol
trialkylglycerol
Cholesteryl oleate
(LDL)
Sphingolipids
Sphingosine
1-O-b-D-galactosyl-sphingosine or
psychosine
N-Acyl-sphingosine or Ceremide
Sphingomyelin
Glycolipids
glycoglycerolipids
glycosphingolipids
cerebrosides
b-D-Galactosyl-diacylglycerol
Common in plants, rare in animals
gangliosides
sialic acid
Lipopolysaccharides
Lipid X
Lipid A
endotoxin
Sterols
H3C
21
12
11
CH3
2
19
10
3
5
1
18
13
9
6
H
25
CH3
26
CH3
27
16
14
8
H
7
23
17
H
H
4
CH3
24
22
20
15
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 = phosphate
neutral or glyco do not stain
Amino Groups (PE and PS)
ninhydrin spray, positive is pink-purple
Glycolipids
a-naphthol give purple spots
Phospholipase Digestions Followed by GC Analysis