Home LiteratureArticle Details
PMID: 10648557 Published · ppublish English Journal Article

nSec1 binds a closed conformation of syntaxin1A.

The Journal of cell biology ·Vol. 148 ·No. 2 ·2000-01-24 ·Pages 247-52

Yang B, Steegmaier M, Gonzalez LC, Scheller RH

Abstract

The Sec1 family of proteins is proposed to function in vesicle trafficking by forming complexes with target membrane SNAREs (soluble N-ethylmaleimide-sensitive factor [NSF] attachment protein [SNAP] receptors) of the syntaxin family. Here, we demonstrate, by using in vitro binding assays, nondenaturing gel electrophoresis, and specific neurotoxin treatment, that the interaction of syntaxin1A with the core SNARE components, SNAP-25 (synaptosome-associated protein of 25 kD) and VAMP2 (vesicle-associated membrane protein 2), precludes the interaction with nSec1 (also called Munc18 and rbSec1). Inversely, association of nSec1 and syntaxin1A prevents assembly of the ternary SNARE complex. Furthermore, using chemical cross-linking of rat brain membranes, we identified nSec1 complexes containing syntaxin1A, but not SNAP-25 or VAMP2. These results support the hypothesis that Sec1 proteins function as syntaxin chaperons during vesicle docking, priming, and membrane fusion.

MeSH Terms
Animals Antigens, Surface/chemistry,genetics,metabolism Biological Transport Botulinum Toxins/pharmacology Membrane Fusion Membrane Proteins/metabolism Molecular Chaperones/genetics,metabolism Munc18 Proteins Nerve Tissue Proteins/chemistry,genetics,metabolism Protein Binding Protein Conformation R-SNARE Proteins Rats Recombinant Proteins/metabolism SNARE Proteins Synaptosomal-Associated Protein 25 Syntaxin 1 Vesicular Transport Proteins
Chemicals
Antigens, Surface Membrane Proteins Molecular Chaperones Munc18 Proteins Nerve Tissue Proteins R-SNARE Proteins Recombinant Proteins SNARE Proteins Snap25 protein, rat Stxbp1 protein, rat Synaptosomal-Associated Protein 25 Syntaxin 1 Vesicular Transport Proteins Botulinum Toxins
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Yang B
Howard Hughes Medical Institute, Department of Molecular Physiology, Stanford University School of Medicine, Stanford, California 94305-5428, USA.
Steegmaier M
Gonzalez L C
Scheller R H
References (27)
27 references, click to expand
  1. Synaptic vesicles and exocytosis.
    Annu Rev Neurosci. 1994;17:219-46 PMID: 8210174
  2. SNARE complex formation is triggered by Ca2+ and drives membrane fusion.
    Cell. 1999 Apr 16;97(2):165-74 PMID: 10219238
  3. Vesicle-associated membrane protein and synaptophysin are associated on the synaptic vesicle.
    J Biol Chem. 1994 Oct 7;269(40):24534-7 PMID: 7929121
  4. rop, a Drosophila homolog of yeast Sec1 and vertebrate n-Sec1/Munc-18 proteins, is a negative regulator of neurotransmitter release in vivo.
    Neuron. 1994 Nov;13(5):1099-108 PMID: 7946348
  5. Sec9 is a SNAP-25-like component of a yeast SNARE complex that may be the effector of Sec4 function in exocytosis.
    Cell. 1994 Oct 21;79(2):245-58 PMID: 7954793
  6. Synaptic vesicle membrane fusion complex: action of clostridial neurotoxins on assembly.
    EMBO J. 1994 Nov 1;13(21):5051-61 PMID: 7957071
  7. Mechanisms of intracellular protein transport.
    Nature. 1994 Nov 3;372(6501):55-63 PMID: 7969419
  8. A molecular description of synaptic vesicle membrane trafficking.
    Annu Rev Biochem. 1994;63:63-100 PMID: 7979250
  9. Sec1p binds to SNARE complexes and concentrates at sites of secretion.
    J Cell Biol. 1999 Jul 26;146(2):333-44 PMID: 10427089
  10. A conformational switch in syntaxin during exocytosis: role of munc18.
    EMBO J. 1999 Aug 16;18(16):4372-82 PMID: 10449403
  11. Secretion and cell-surface growth are blocked in a temperature-sensitive mutant of Saccharomyces cerevisiae.
    Proc Natl Acad Sci U S A. 1979 Apr;76(4):1858-62 PMID: 377286
  12. The yeast SLY gene products, suppressors of defects in the essential GTP-binding Ypt1 protein, may act in endoplasmic reticulum-to-Golgi transport.
    Mol Cell Biol. 1991 Jun;11(6):2980-93 PMID: 1903839
  13. Synaptic vesicle fusion complex contains unc-18 homologue bound to syntaxin.
    Nature. 1993 Nov 25;366(6453):347-51 PMID: 8247129
  14. n-Sec1: a neural-specific syntaxin-binding protein.
    Proc Natl Acad Sci U S A. 1994 Feb 15;91(4):1445-9 PMID: 8108429
  15. Protein-protein interactions contributing to the specificity of intracellular vesicular trafficking.
    Science. 1994 Feb 25;263(5150):1146-9 PMID: 8108733
  16. In chromaffin cells, the mammalian Sec1p homologue is a syntaxin 1A-binding protein associated with chromaffin granules.
    J Biol Chem. 1994 Mar 25;269(12):8623-6 PMID: 8132588
  17. A rat brain Sec1 homologue related to Rop and UNC18 interacts with syntaxin.
    Proc Natl Acad Sci U S A. 1994 Mar 15;91(6):2003-7 PMID: 8134339
  18. Mutations in the VPS45 gene, a SEC1 homologue, result in vacuolar protein sorting defects and accumulation of membrane vesicles.
    J Cell Sci. 1994 Dec;107 ( Pt 12):3449-59 PMID: 7706396
  19. SNAREs and the specificity of transport vesicle targeting.
    Curr Opin Cell Biol. 1995 Aug;7(4):581-6 PMID: 7495579
  20. ROP, the Drosophila Sec1 homolog, interacts with syntaxin and regulates neurotransmitter release in a dosage-dependent manner.
    EMBO J. 1998 Jan 2;17(1):127-39 PMID: 9427747
  21. SNAREpins: minimal machinery for membrane fusion.
    Cell. 1998 Mar 20;92(6):759-72 PMID: 9529252
  22. The synaptic SNARE complex is a parallel four-stranded helical bundle.
    Nat Struct Biol. 1998 Sep;5(9):765-9 PMID: 9731768
  23. Three-dimensional structure of an evolutionarily conserved N-terminal domain of syntaxin 1A.
    Cell. 1998 Sep 18;94(6):841-9 PMID: 9753330
  24. Crystal structure of a SNARE complex involved in synaptic exocytosis at 2.4 A resolution.
    Nature. 1998 Sep 24;395(6700):347-53 PMID: 9759724
  25. Vac1p coordinates Rab and phosphatidylinositol 3-kinase signaling in Vps45p-dependent vesicle docking/fusion at the endosome.
    Curr Biol. 1999 Feb 11;9(3):159-62 PMID: 10021387
  26. SNARE interactions are not selective. Implications for membrane fusion specificity.
    J Biol Chem. 1999 Feb 26;274(9):5649-53 PMID: 10026182
  27. Specificity and regulation of a synaptic vesicle docking complex.
    Neuron. 1994 Aug;13(2):353-61 PMID: 8060616
Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.148.2.247
Published
2000-01-24
Pages
247-52
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2174276
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.148.2.247 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2