Day 10 MB Lipids

• Monday, Sept. 7: No Class – Labor Day After reading the text, attending lecture, and
• Wednesday, Sept. 9: Membrane lipids, Ch. 10 reviewing lecture notes, you should be
– Quiz on Week 3 due by 1 PM able to
• Friday, Sept. 11: Membrane proteins, Ch. 10 • Explain the significance of asymmetry of
• Monday, Sept. 14: Membrane transport 1, Ch. 11 membrane monolayers.
– Exam 1 on Days 1-10 due by 1 PM • Describe experimental evidence for
membrane lipid fluidity.
I. Membrane lipids
• Describe different types of membrane
A. Quiz Review
proteins.
B. Fluid mosaic model
C. Membrane asymmetry • Perform hydropathy analysis on a protein
D. Evidence for membrane fluidity sequence to predict transmembrane
regions.
II. Membrane proteins
A. RBC MB proteins
B. Classes
C. Hydropathy analysis
B. Fluid Mosaic Model
Singer and Nicholson (1972) Science 175:720
C. Asymmetry and Movement of PLs
• Functional significance:
• Contributes to net negative charge inside
• PI is available for signaling function inside.
• Glycolipids in outer leaflet, so most CHO
out.
• Recognition of dead cells

• Inequality is maintained by movement

properties of phospholipids within the
membrane
• In plasma membrane: – Rotation and lateral diffusion is rapid
• PC, SM mostly in outer leaflet
– Transverse diffusion or "flip-flop" is rare
• PE, PS, PI mostly in inner leaflet
• Cholesterol: 50% inner, 50% outer
• How do they get that way? Enzymes move
them after membrane synthesis in the ER.
D. Lipid Fluidity: Evidence from
Fluorescence Recovery After Photobleaching (FRAP)

Lipids
labeled
Evidence for Fluidity:
Differential Scanning Calorimetry
• Measures absorption of heat during phase
transitions of lipids.

• Below the transition temperature (Tm) lipids are

solid, above Tm lipids are fluid.

• Longer fatty acid chains have a higher Tm while

shorter fatty acids have a lower Tm (more fluid).

• Saturated fatty acids have a higher Tm while

unsaturated fatty acids have a lower Tm (more
fluid). Why?
Monoun-
saturated
saturated
MB Fluidity Depends On:
• Temperature
– Higher T, greater fluidity; cells can’t change.
• Unsaturated FAs
– Increase fluidity
• Length of FAs
– Shorter, more fluid Cells can regulate
• Cholesterol
– Fluidity “buffer”
Effect of Unsaturated Fatty Acids on Fluidity
• C=C in FA creates kinks in
chain, so they pack together
less well.

• Less able to form crystalline

solid, therefore stays liquid.

• Organisms in cold
environments increase the #
of unsaturated FAs in their
membranes.
Effects of Chain Length and Double Bonds on Tm

Less fluid → More fluid →

Buffering Effect of Cholesterol on Fluidity
• Animal cells contain up to 50% cholesterol in
their membranes.
• OH of cholesterol hydrogen bonds with O of
ester bonded fatty acid, while hydrocarbon
rings interact with hydrophobic hydrocarbon
chains of FAs

Acts as a fluidity buffer:

Makes MB less fluid at higher
temperatures than without
cholesterol, since FAs less mobile.
Makes MB more fluid at lower
temperatures than without
cholesterol, since it disrupts packing
into a crystal.
A recent twist on the Fluid Mosaic Model: Lipid rafts

Or
Outside cell

• Small, specialized areas in membrane where some lipids (LC FA’s, sphingolipids and cholesterol)
and proteins are concentrated.
– Two monolayers in raft move together; thicker, less fluid than normal membrane
• Function: concentrating membrane proteins for vesicle transport and/or coordination of
pathways.
 Summary: Evidence concerning the Lipid
Portion of the Membrane
1. Estimated and measured surface area
• Membrane is a bilayer.

2. Electron microscopy
• Trilaminar appearance of membranes.

3. Thin-layer chromatography
• Different membranes contain different phospholipids.

4. Fluorescence recovery after photobleaching of lipids

• Membranes are fluid.

5. Differential scanning calorimetry

• The lipid composition of membranes determines how fluid they are.
II. Membrane Proteins
A. RBC membrane proteins: PAGE and diagram

From: Harper’s Illustrated Biochemistry (2012)

http://schoolbag.info/chemistry/biochemistry_1/58.html
An underlying cortex
reinforces the
plasma membrane
(anchors)
B. Classes of Membrane Proteins

1. Integral membrane proteins (A-C): require detergent to remove from MB

2. Peripheral membrane proteins (D): removed by milder treatments (high salt, ∆pH or >[Ca 2+ ])
since attached by weaker non-covalent interactions
Q: What effect on a gel would you see if each of the above was treated with protease on the
outside?
Some membrane proteins attach to lipids
C. Hydropathy analysis to ID transmembrane
segments
Bacteriorhodopsin
• One of first TM proteins whose
structure was determined.

• 7-pass transmembrane protein.

• Transmembrane regions are  -helical

with hydrophobic R groups facing out -
usually 20-30 amino acids.

• Functions as a light-driven proton pump

in membrane of certain Archaea.

• Creates proton gradient across PM used

to make ATP.