SNARE Interactions Are Not Selective
IMPLICATIONS FOR MEMBRANE FUSION SPECIFICITY*
- Bin Yang‡,
- Lino Gonzalez, Jr.‡§,
- Rytis Prekeris,
- Martin Steegmaier,
- Raj J. Advani and
- Richard H. Scheller¶
- From the Howard Hughes Medical Institute, Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, California 94305-5428
Abstract
The SNARE hypothesis proposes that membrane trafficking specificity is mediated by preferential high affinity interactions between particular v (vesicle membrane)- and t (target membrane)-SNARE combinations. The specificity of interactions among a diverse set of SNAREs, however, is unknown. We have tested the SNARE hypothesis by analyzing potential SNARE complexes between five proteins of the vesicle-associated membrane protein (VAMP) family, three members of the synaptosome-associated protein-25 (SNAP-25) family and three members of the syntaxin family. All of the 21 combinations of SNAREs tested formed stable complexes. Sixteen were resistant to SDS denaturation, and most complexes thermally denatured between 70 and 90 °C. These results suggest that the specificity of membrane fusion is not encoded by the interactions between SNAREs.
Footnotes
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↵* 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.
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↵‡ These authors contributed equally to this work.
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↵§ Supported by a National Science Foundation postdoctoral fellowship awarded in 1997.
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↵¶ Investigator of the Howard Hughes Medical Institute. To whom correspondence should be addressed. Tel.: 650-723-9075; Fax: 650-725-4436; E-mail: scheller{at}cmgm.stanford.edu.
- Abbreviations:
- SNARE
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SNAP receptor
- SNAP
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soluble NSF attachment protein
- NSF
-
N-ethylmaleimide-sensitive factor
- SNAP-23
-
-25, -29, synaptosome-associated protein of 23, 25, and 29 kDa
- r-SNARE
-
arginine class of SNARE
- q-SNARE
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glutamine class of SNARE
- VAMP
-
vesicle-associated membrane protein
- PAGE
-
polyacrylamide gel electrophoresis
- GST
-
glutathione S-transferase
- PCR
-
polymerase chain reaction
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- Received December 23, 1998.
- The American Society for Biochemistry and Molecular Biology, Inc.
