A fundamental question in cell biology is how the recruitment of proteins to cellular membranes is achieved and regulated. The long term objective of the proposed research is to characterize the structural and energetic basis for the binding of peripheral membrane proteins to phospholipid membranes and, in turn, to better understand the biophysical basis of membrane-mediated events in cells. The overall strategy is based on two distinct computational approaches: 1) the calculation of the physical interactions between proteins and membranes, and 2) bioinformatics tools for sequence analysis, structure comparison and structure prediction. By studying different membrane-interacting proteins in parallel, two hypotheses will be tested: 1) that two physical factors-electrostatics and hydrophobicity-are the major determinants of membrane binding, and 2) that these physical factors are manifested as patterns in sequence, structure and biophysical characteristics that can be used to predict membrane targeting potential. The first specific aim is to describe how nonspecific electrostatic and hydrophobic interactions mediate the wide range of membrane binding behaviors exhibited by secreted phospholipases A2. The second specific aim is to understand the role of electrostatic interactions in the calcium-dependent and independent membrane binding of C2 domains. The third specific aim is to develop structural models for phosphoinositides, an important class of signaling lipids. The fourth specific aim is to determine the energetic basis of both the specific and non-specific interactions of pleckstrin homology (PH) domains with membranes containing phosphoinositides. The computational results will be used in the design and interpretation of experiments through collaborations with experimental groups and will lead to rules that can be used to detect membrane targeting motifs in proteins. Secreted phospholipases A2 have been implicated in inflammation, and C2 and PH domain-containing proteins involved in phosphoinositide signaling have been implicated in oncogenesis; these proteins require membrane association for their function. Thus, a detailed understanding of the molecular mechanisms underlying the control of membrane association would facilitate the rational design of drugs that inhibit membrane binding.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM066147-04
Application #
6932493
Study Section
Molecular and Cellular Biophysics Study Section (BBCA)
Program Officer
Chin, Jean
Project Start
2002-08-15
Project End
2007-07-31
Budget Start
2005-08-01
Budget End
2006-07-31
Support Year
4
Fiscal Year
2005
Total Cost
$245,630
Indirect Cost
Name
Weill Medical College of Cornell University
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
060217502
City
New York
State
NY
Country
United States
Zip Code
10065
Li, Zheng; Venable, Richard M; Rogers, Laura A et al. (2009) Molecular dynamics simulations of PIP2 and PIP3 in lipid bilayers: determination of ring orientation, and the effects of surface roughness on a Poisson-Boltzmann description. Biophys J 97:155-63
Blatner, Nichole R; Wilson, Michael I; Lei, Cai et al. (2007) The structural basis of novel endosome anchoring activity of KIF16B kinesin. EMBO J 26:3709-19
Dalton, Amanda K; Ako-Adjei, Danso; Murray, Paul S et al. (2007) Electrostatic interactions drive membrane association of the human immunodeficiency virus type 1 Gag MA domain. J Virol 81:6434-45
Mulgrew-Nesbitt, Anna; Diraviyam, Karthikeyan; Wang, Jiyao et al. (2006) The role of electrostatics in protein-membrane interactions. Biochim Biophys Acta 1761:812-26
Wijewickrama, Gihani T; Albanese, Alexandra; Kim, Young Jun et al. (2006) Unique membrane interaction mode of group IIF phospholipase A2. J Biol Chem 281:32741-54
Diraviyam, Karthikeyan; Murray, Diana (2006) Computational analysis of the membrane association of group IIA secreted phospholipases A2: a differential role for electrostatics. Biochemistry 45:2584-98
Evans, John H; Murray, Diana; Leslie, Christina C et al. (2006) Specific translocation of protein kinase Calpha to the plasma membrane requires both Ca2+ and PIP2 recognition by its C2 domain. Mol Biol Cell 17:56-66
Stahelin, Robert V; Wang, Jiyao; Blatner, Nichole R et al. (2005) The origin of C1A-C2 interdomain interactions in protein kinase Calpha. J Biol Chem 280:36452-63
Bollinger, James G; Diraviyam, Karthikeyan; Ghomashchi, Farideh et al. (2004) Interfacial binding of bee venom secreted phospholipase A2 to membranes occurs predominantly by a nonelectrostatic mechanism. Biochemistry 43:13293-304
Evans, John H; Gerber, Stefan H; Murray, Diana et al. (2004) The calcium binding loops of the cytosolic phospholipase A2 C2 domain specify targeting to Golgi and ER in live cells. Mol Biol Cell 15:371-83

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