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Originally published In Press as doi:10.1074/jbc.M101513200 on March 12, 2001

J. Biol. Chem., Vol. 276, Issue 21, 18178-18184, May 25, 2001
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Sequential Involvement of p115, SNAREs, and Rab Proteins in Intra-Golgi Protein Transport*

Michael J. S. GmachlDagger and Christian Wimmer§

From the Dagger  Institute for Molecular Pathology, Dr. Bohrgasse 7, A-1030 Vienna, Austria and the § Basel Institute for Immunology, Grenzacherstraße 487, Ch-4005 Basel, Switzerland

Delivery of transport vesicles to their receptor compartment involves tethering, priming, and fusion. Soluble NSF attachment protein-alpha (alpha SNAP) mediates the disruption of SNAREs by N-ethylmaleimide sensitive factor (NSF) and was employed to determine the hierarchy of proteins responsible for intra-Golgi protein transport. The N-terminal 23 amino acids of alpha SNAP are necessary for SNARE binding. The antibody 2F10 recognizes this SNARE interaction domain of alpha SNAP and inhibits intra-Golgi protein transport reversibly. This antibody was applied to modify the transport assay to determine the protein requirements relative to the action of alpha SNAP and NSF. We found that 1) p115 acts independently of alpha SNAP and NSF, 2) SNAREs are required after tethering and interact selectively after activation by alpha SNAP and NSF, and 3) Rab proteins act after SNARE activation and before fusion.

* This work was supported by a postdoctoral fellowship from the Max Kade Foundation (to M. J. S. G.) and a postdoctoral fellowship from the Deutsche Forschungsgemeinschaft (to C. W.). The Basel Institute for Immunology was founded and is supported by F. Hoffmann-La Roche Ltd., Basel, Switzerland.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

To whom correspondence should be addressed. Tel.: 41616051351; Fax: 41616051364; E-mail: wimmer@bii.ch.

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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This article has been cited by other articles:


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K. E. Marz, J. M. Lauer, and P. I. Hanson
Defining the SNARE Complex Binding Surface of {alpha}-SNAP: IMPLICATIONS FOR SNARE COMPLEX DISASSEMBLY
J. Biol. Chem., July 11, 2003; 278(29): 27000 - 27008.
[Abstract] [Full Text] [PDF]


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