The long-term goals of this project are to develop methods to probe the organization and dynamic reorganization of biological membranes. This includes interactions among the components within membranes, interactions between membrane surfaces that lead to binding, fusion, and pattern formation, and conformational changes of proteins associated with membranes. The lipid bilayer is the basic structure common to biological membranes. Membrane fluidity is critical for biological functions that depend upon conformational changes within membranes, the lateral association or clustering of multiple components, and processes that change membrane topology such as edo- and exocytocis and fusion. This proposal outlines new types of experiments that probe these basic aspects of membrane dynamics using tools that have been developed to pattern, manipulate and image supported bilayers. During the next grant period the focus will be on the mechanism of vesicle fusion, using vesicles that are tethered to supported bilayers and whose interactions can be monitored at the level of individual vesicles (Aim 1);the lateral association and organization of lipids and membrane anchored proteins using a novel type of imaging mass spectrometry that permits membrane composition analysis with unprecedented lateral resolution, sensitivity and information content (Aim 2);and the design and fabrication of an integrated optical/electrical device that will permit high precision interferometry on planar bilayers to probe conformational transitions of membrane-associated proteins, with an initial focus on voltage-gated ion channels (Aim 3).
Each aim depends upon the development of new supported lipid bilayer architectures and analytical methods that can have a broad impact on studies of biological membranes. Relevance to human health: A significant fraction of all proteins are associated with membranes, and, as a class, these constitute a huge and diverse target for drug development. This proposal outlines new methods for studying membranes and membrane-associated proteins that can impact our understanding of biological function and organization, as well as impact biotechnology.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM069630-07
Application #
7753656
Study Section
Biochemistry and Biophysics of Membranes Study Section (BBM)
Program Officer
Chin, Jean
Project Start
2004-01-01
Project End
2011-12-31
Budget Start
2010-01-01
Budget End
2010-12-31
Support Year
7
Fiscal Year
2010
Total Cost
$375,078
Indirect Cost
Name
Stanford University
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
009214214
City
Stanford
State
CA
Country
United States
Zip Code
94305
Flavier, Kristina M; Boxer, Steven G (2017) Vesicle Fusion Mediated by Solanesol-Anchored DNA. Biophys J 113:1260-1268
Fried, Stephen D; Boxer, Steven G (2017) Electric Fields and Enzyme Catalysis. Annu Rev Biochem 86:387-415
Lozano, Mónica M; Hovis, Jennifer S; Moss 3rd, Frank R et al. (2016) Dynamic Reorganization and Correlation among Lipid Raft Components. J Am Chem Soc 138:9996-10001
Mercer, Jaron A M; Cohen, Carolyn M; Shuken, Steven R et al. (2016) Chemical Synthesis and Self-Assembly of a Ladderane Phospholipid. J Am Chem Soc 138:15845-15848
Moss 3rd, Frank R; Boxer, Steven G (2016) Atomic Recombination in Dynamic Secondary Ion Mass Spectrometry Probes Distance in Lipid Assemblies: A Nanometer Chemical Ruler. J Am Chem Soc 138:16737-16744
Rawle, Robert J; Boxer, Steven G; Kasson, Peter M (2016) Disentangling Viral Membrane Fusion from Receptor Binding Using Synthetic DNA-Lipid Conjugates. Biophys J 111:123-31
Hughes, Laura D; Rawle, Robert J; Boxer, Steven G (2014) Choose your label wisely: water-soluble fluorophores often interact with lipid bilayers. PLoS One 9:e87649
van Lengerich, Bettina; Rawle, Robert J; Bendix, Poul Martin et al. (2013) Individual vesicle fusion events mediated by lipid-anchored DNA. Biophys J 105:409-19
Hughes, Laura D; Boxer, Steven G (2013) DNA-based patterning of tethered membrane patches. Langmuir 29:12220-7
Chung, Minsub; Koo, Bon Jun; Boxer, Steven G (2013) Formation and analysis of topographical domains between lipid membranes tethered by DNA hybrids of different lengths. Faraday Discuss 161:333-45; discussion 419-59

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