Abstract

The aim of the proposed research is to develop newly designed membrane mimetics for NMR studies of the structure and dynamics of membrane associated proteins and peptides. These mimetics consist of detergent free mixtures of naturally occurring phospholipids with both neutral and anionic headgroups. The mixtures have a versatile phase diagram that includes an isotropic region as well as a magnetically ordered phase that can have either a positive or negative order parameter. In this proposal, the value of these mimetics will be demonstrated by carrying out high resolution and solid state NMR studies of representative membrane bound peptides and proteins chosen to illustrate the wide range of unsolved structural problems. Examples include, peptides from the myristoylated N-terminal segments of cAMP dependent protein kinase and a src tyrosine kinase, orphanin FQ (a neuropeptide), phospholipase A2, bombolin (a PLA2 activator) and the membrane spanning segment of glycophorin.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM054034-02
Application #
2701727
Study Section
Biophysical Chemistry Study Section (BBCB)
Project Start
1997-05-01
Project End
2000-04-30
Budget Start
1998-05-01
Budget End
1999-04-30
Support Year
2
Fiscal Year
1998
Total Cost
Indirect Cost

  Institution

Name
University of California San Diego
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
077758407
City
La Jolla
State
CA
Country
United States
Zip Code
92093

  Publications

Whiles, Jennifer A; Glover, Kerney J; Vold, Regitze R et al. (2002) Methods for studying transmembrane peptides in bicelles: consequences of hydrophobic mismatch and peptide sequence. J Magn Reson 158:149-56
Whiles, Jennifer A; Deems, Raymond; Vold, Regitze R et al. (2002) Bicelles in structure-function studies of membrane-associated proteins. Bioorg Chem 30:431-42
Glover, Kerney Jebrell; Whiles, Jennifer A; Vold, Regitze R et al. (2002) Position of residues in transmembrane peptides with respect to the lipid bilayer: a combined lipid Noes and water chemical exchange approach in phospholipid bicelles. J Biomol NMR 22:57-64
Whiles, J A; Brasseur, R; Glover, K J et al. (2001) Orientation and effects of mastoparan X on phospholipid bicelles. Biophys J 80:280-93
Glover, K J; Whiles, J A; Wood, M J et al. (2001) Conformational dimorphism and transmembrane orientation of prion protein residues 110-136 in bicelles. Biochemistry 40:13137-42
Glover, K J; Whiles, J A; Wu, G et al. (2001) Structural evaluation of phospholipid bicelles for solution-state studies of membrane-associated biomolecules. Biophys J 81:2163-71
Struppe, J; Whiles, J A; Vold, R R (2000) Acidic phospholipid bicelles: a versatile model membrane system. Biophys J 78:281-9
Struppe, J; Vold, R R (1998) Dilute bicellar solutions for structural NMR work. J Magn Reson 135:541-6
Struppe, J; Komives, E A; Taylor, S S et al. (1998) 2H NMR studies of a myristoylated peptide in neutral and acidic phospholipid bicelles. Biochemistry 37:15523-7