A deep understanding of ttie physiology and biophysics of fusion - and its disease relevance - requires a broad and comprehensive approach and relies heavily upon interdisciplinary approaches honed from a decade of studies of the mechanism of viral envelope fusion. These technologies - cell and molecular biology, surface force/adhesion biophysics, and optical imaging.
The specific aims ofthe initial application covered four aspects of SNARE induced fusion: 1) SNARE distribution, mobility, and lateral interactions within the plane ofthe membrane. 2) Adhesive and molecular forces between SNAREs - determinants of transition states and the energetic. 3) Penultimate states of SNARE fusion: capturing the SNAREpin just before lipid-mixing, with a specific focus on the interplay between lipid structure and protein assembly. 4) Protein and lipidic determinants of fusion kinetics and pore formation.

Public Health Relevance

This broad and comprehensive approach to elucidating the mechanism of intracellular membrane fusion by SNARE proteins will deepen our understanding ofthe physiology and biophysics effusion - and its key roles in diabetes.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
5R37DK027044-37
Application #
8280410
Study Section
Special Emphasis Panel (NSS)
Program Officer
Haft, Carol R
Project Start
1991-09-30
Project End
2014-06-30
Budget Start
2012-07-01
Budget End
2014-06-30
Support Year
37
Fiscal Year
2012
Total Cost
$671,580
Indirect Cost
$253,057
Name
Yale University
Department
Anatomy/Cell Biology
Type
Schools of Medicine
DUNS #
043207562
City
New Haven
State
CT
Country
United States
Zip Code
06520
Bello, Oscar D; Auclair, Sarah M; Rothman, James E et al. (2016) Using ApoE Nanolipoprotein Particles To Analyze SNARE-Induced Fusion Pores. Langmuir 32:3015-23
Xu, Weiming; Nathwani, Bhavik; Lin, Chenxiang et al. (2016) A Programmable DNA Origami Platform to Organize SNAREs for Membrane Fusion. J Am Chem Soc 138:4439-47
Li, Feng; Tiwari, Neeraj; Rothman, James E et al. (2016) Kinetic barriers to SNAREpin assembly in the regulation of membrane docking/priming and fusion. Proc Natl Acad Sci U S A 113:10536-41
Xu, Weiming; Wang, Jing; Rothman, James E et al. (2015) Accelerating SNARE-Mediated Membrane Fusion by DNA-Lipid Tethers. Angew Chem Int Ed Engl 54:14388-92
Li, Feng; Kümmel, Daniel; Coleman, Jeff et al. (2014) A half-zippered SNARE complex represents a functional intermediate in membrane fusion. J Am Chem Soc 136:3456-64
Zorman, Sylvain; Rebane, Aleksander A; Ma, Lu et al. (2014) Common intermediates and kinetics, but different energetics, in the assembly of SNARE proteins. Elife 3:e03348
Shi, Lei; Howan, Kevin; Shen, Qing-Tao et al. (2013) Preparation and characterization of SNARE-containing nanodiscs and direct study of cargo release through fusion pores. Nat Protoc 8:935-48
Shi, Lei; Shen, Qing-Tao; Kiel, Alexander et al. (2012) SNARE proteins: one to fuse and three to keep the nascent fusion pore open. Science 335:1355-9
Kümmel, Daniel; Krishnakumar, Shyam S; Radoff, Daniel T et al. (2011) Complexin cross-links prefusion SNAREs into a zigzag array. Nat Struct Mol Biol 18:927-33
Shen, Jingshi; Rathore, Shailendra S; Khandan, Lavan et al. (2010) SNARE bundle and syntaxin N-peptide constitute a minimal complement for Munc18-1 activation of membrane fusion. J Cell Biol 190:55-63

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