1. Hydrophobic Mismatch Lateral Pressure These lecture notes are taken from current literature. See special edition of BBA, (biomembranes) 1666 2004.

2. Our previous lectures have discussed in detail what is meant by “non- bilayer” structures (really non-lamellar because inverted hexagonal phase is somewhat bilayer in nature). This figure reviews how shape is related to mesophase.

3. Effects of “non-bilayer” lipids on membrane proteins: Curvature Stress – discussed in detail in previous lectures Hydrophobic mismatch Packing Defects Lateral Pressure Profiles Our Motivation: To understand how lipids and lipid-protein interactions drive membrane protein function and membrane-protein interactions Different types of “membrane” proteins

4. Hydrophobic Mismatch When the hydrophobic region of a protein or peptide (length) differs from that of the bilayer, the bilayer will adapt and curve to “cover” the hydrophobic surface area of the protein. Sometimes the protein will also change it’s conformation or tilt. Changes in bilayer shape are related to both the curvature near the protein as well as “global” lipid bilayer thinning or thickening. The WALP peptide are the classic study of effects of hydrophobic mismatch both for membrane properties and peptide tilt. WALP (ALA and LEU with TRP to anchor at the bilayer interface)

5. Protein Sorting Conformational Changes Rafts and Lateral organization Hydrophobic Mismatch

6. Hydrophobic Mismatch: Protein Folding and Sorting

7. Hydrophobic Mismatch Effects: Gramicidin and Rhodopsin Extensively Studied VpU Tilt of protein can change as a function of membrane thickness Oligomerization states can change

8. Conformations can change Hydrophobic Mismatch Question: How do non-lamellar lipids affect hydrophobic mismatch?

9. Packing Defects/Insertion Sites Several studies have suggested that the presence of the non-lamellar lipids will produce bilayer packing defects that can influence peripheral membrane proteins. Asymmetry can produce a physical curvature Symmetric distribution of non-lamellar lipid Examples of protein binding effected by lipids with negative spontaneous Curvature: phospholipase A2 Protein kinase C Apolopophirin II CTP:phosphocholine cytidylyltransferase

10. Lateral Pressure In reality, the spontaneous curvature, defect packing and lateral pressure concepts are related. Lateral Pressure Profile in membranes. First proposed in 1960s. Large negative pressure due to the cohesive hydrophobic interfacial tension is localized at the polar-apolar interface

11. Overall Lateral Pressure is zero (integration over profile must be zero) There curvature stress is related to the lateral pressure profile via the torque tension, , which is the first moment of the lateral pressure. Remember, spontaneous curvature is not real curvature, it is a representation about stored energy/forces in the bilayer. Forces can be related to pressures. Lateral pressure Spontaneous curvature Lateral Pressure

12. Changes in the lateral pressure profile will only affect those membrane proteins whose function involves a conformational change that is accompanied by a depth dependent variation in cross- sectional area. Lateral Pressure

13. Examples of Proteins Affected by Lateral pressure profiles: leader peptidase  2-75 KcsA

14. KcsA – symmetric tetramer with each monomer consisting of an N-terminal helix at the membrane interface, a transmembrane helix, followed by a short pore helix leading to the selectivity filter in the tetrameric structure The KcsA tetramer is stable in SDS, but can be denatured by heat or TFE-trifluoroethanol. KcsA preferentially interacts with PE and PG lipids. Tetramer stability: detergent < cylindrical PC < PC/PG (7:3) < PE/PG (7:3) TFE interactions with lipid bilayers interact with head group region, decreasing the acylchain order

15. DOPCDOPG/DOPE DOPC PG/PE Concept: an “hour- glass” shaped oligomer is stabilized in bilayer mixtures with PE. Addition of TFE perturbs the lateral pressure profiles