Membrane fusion at neuronal synapses is a highly regulated process that is triggered by micromolar concentrations of calcium in the presynaptic cell. The SNARE proteins drive fusion and form the critical core of the fusion machinery;however, they are not directly regulated by calcium. Synaptotagmin 1 is a vesicle associated membrane protein that functions as the calcium sensor in neuronal exocytosis. It is anchored by a single transmembrane segment at its N-terminus and contains two C2 domains that have been shown to bind membranes in a calcium-dependent fashion. Synaptotagmin also interacts with SNAREs. At the present time, the nature of the synaptotagmin interactions that mediate fusion are not understood. The proposed work will utilize evolving magnetic resonance methods, such as site-directed spin labeling and pulse EPR, to evaluate a number of proposed mechanisms for the molecular function of sytnaptotagmin.
The specific aims are to test the importance of a bridging conformation that is observed for synaptotagmin 1, where the two C2 domains bind opposing bilayers. Proposed experiments will test the hypothesis that synaptotagmin 1 switches from SNARE to membrane interactions in the presence of calcium, and both membrane and SNARE associated structures for synaptotagmin 1 will be generated.
A final aim will examine the effects of synaptotagmin 1 on lipid bilayer properties and test the role of these effects in mediating membrane fusion. Membrane fusion is central to cellular processes such as intracellular transport, host defense (killing of microorganisms, immune response), and human physiology and disease (e.g., glucose regulation/diabetes, allergic response). Calcium -triggered neuronal fusion is a central process in neurobiology and it underlies synaptic transmission and potentiation. Regulated neuronal exocytosis is thought to be relevant to an understanding of schizophrenia, bipolar disorder, Huntington's disease, Parkinson's disease, Alzheimer's disease as well as range of other neurological disorders.

Public Health Relevance

The proposed work is directed at characterizing the molecular mechanisms by which neurons release neurotransmitters and communicate with each other in the central nervous system. The mechanisms of neurotransmitter release are relevant to understanding a wide range of neurological disorders, including schizophrenia, bipolar disorder, Huntington's disease, Parkinson's disease and Alzheimer's disease.

Agency
National Institute of Health (NIH)
Type
Research Program Projects (P01)
Project #
5P01GM072694-09
Application #
8706172
Study Section
Special Emphasis Panel (ZRG1)
Project Start
Project End
Budget Start
Budget End
Support Year
9
Fiscal Year
2014
Total Cost
Indirect Cost
Name
University of Virginia
Department
Type
DUNS #
City
Charlottesville
State
VA
Country
United States
Zip Code
22904
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Kiessling, Volker; Yang, Sung-Tae; Tamm, Lukas K (2015) Supported lipid bilayers as models for studying membrane domains. Curr Top Membr 75:1-23
Kreutzberger, Alex J B; Kiessling, Volker; Tamm, Lukas K (2015) High cholesterol obviates a prolonged hemifusion intermediate in fast SNARE-mediated membrane fusion. Biophys J 109:319-29
Milovanovic, Dragomir; Honigmann, Alf; Koike, Seiichi et al. (2015) Hydrophobic mismatch sorts SNARE proteins into distinct membrane domains. Nat Commun 6:5984

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