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Structural Analysis of the Neuronal SNARE Protein Syntaxin-1A,

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Department of Molecular Biology, Princeton University, Princeton, New Jersey 08544, and Department of Cell, Molecular, and Developmental Biology, Haverford College, Haverford, Pennsylvania 19041
Cite this: Biochemistry 2000, 39, 29, 8470–8479
Publication Date (Web):June 23, 2000
Copyright © 2000 American Chemical Society

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    Intracellular trafficking depends on the docking and fusion of transport vesicles with cellular membranes. Central to docking and fusion is the pairing of SNARE proteins (soluble NSF attachment protein receptors) associated with the vesicle and target membranes (v- and t-SNAREs, respectively). Here, the X-ray structure of an N-terminal conserved domain of the neuronal t-SNARE syntaxin-1A was determined to a resolution of 1.9 Å using multiwavelength anomalous diffraction. This X-ray structure, which is in general agreement with an NMR structure of a similar fragment, provides new insight into the interaction surface between the N-terminal domain and the remainder of the protein. In vitro characterization of the intact cytoplasmic domain of syntaxin revealed that it forms dimers, and probably tetramers, at low micromolar concentrations, with concomitant structural changes that can be detected by limited proteolysis. These observations suggest that the promiscuity characteristic of pairing between v-SNAREs and t-SNAREs extends to the formation of homo-oligomeric t-SNARE complexes as well. They also suggest a potential role for the neuronal Sec1 protein (nSec1) in preventing the formation of syntaxin multimers.

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     This work was supported by NIH Grant NS-38046, the Searle Scholars Program, and the Beckman Young Investigators Program (F.M.H.).

     X-ray coordinates and structure factors have been deposited in the Research Collaboratory for Structural Bioinformatics, Rutgers University, New Brunswick, NJ (1EZ3).


     Princeton University.

     Haverford College.


     To whom correspondence should be addressed. Telephone:  (609) 258-4982. Fax:  (609) 258-6730. E-mail:  [email protected].

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