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PMID: 22508512 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Munc13-4 reconstitutes calcium-dependent SNARE-mediated membrane fusion.

The Journal of cell biology ·Vol. 197 ·No. 2 ·2012-04-16 ·Pages 301-12

Boswell KL, James DJ, Esquibel JM, Bruinsma S, Shirakawa R, Horiuchi H, Martin TF

Abstract

Munc13-4 is a widely expressed member of the CAPS/Munc13 protein family proposed to function in priming secretory granules for exocytosis. Munc13-4 contains N- and C-terminal C2 domains (C2A and C2B) predicted to bind Ca(2+), but Ca(2+)-dependent regulation of Munc13-4 activity has not been described. The C2 domains bracket a predicted SNARE-binding domain, but whether Munc13-4 interacts with SNARE proteins is unknown. We report that Munc13-4 bound Ca(2+) and restored Ca(2+)-dependent granule exocytosis to permeable cells (platelets, mast, and neuroendocrine cells) dependent on putative Ca(2+)-binding residues in C2A and C2B. Munc13-4 exhibited Ca(2+)-stimulated SNARE interactions dependent on C2A and Ca(2+)-dependent membrane binding dependent on C2B. In an apparent coupling of membrane and SNARE binding, Munc13-4 stimulated SNARE-dependent liposome fusion dependent on putative Ca(2+)-binding residues in both C2A and C2B domains. Munc13-4 is the first priming factor shown to promote Ca(2+)-dependent SNARE complex formation and SNARE-mediated liposome fusion. These properties of Munc13-4 suggest its function as a Ca(2+) sensor at rate-limiting priming steps in granule exocytosis.

MeSH Terms
Blood Platelets/metabolism Calcium/metabolism Calcium-Binding Proteins/metabolism Exocytosis/physiology Humans Liposomes/metabolism Mast Cells/metabolism Membrane Fusion/physiology Membrane Proteins/metabolism Neuroendocrine Cells/metabolism SNARE Proteins/metabolism Synaptotagmins/metabolism
Chemicals
Calcium-Binding Proteins Liposomes Membrane Proteins SNARE Proteins UNC13D protein, human Synaptotagmins Calcium
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Boswell Kristin L
Department of Biochemistry, University of Wisconsin, Madison, WI 53706, USA.
James Declan J
Esquibel Joseph M
Bruinsma Stephen
Shirakawa Ryutaro
Horiuchi Hisanori
Martin Thomas F J
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
1540-8140
DOI
10.1083/jcb.201109132
Published
2012-04-16
Pages
301-12
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC3328385
Subset
IM
Grants
NIGMS NIH HHS · R01 GM119158 · United States
NIDDK NIH HHS · R37 DK025861 · United States
NIDDK NIH HHS · R01 DK025861 · United States
NIGMS NIH HHS · GM08668 · United States
NIDDK NIH HHS · DK025861 · United States
NIDDK NIH HHS · R01 DK040428 · United States
External Links
PubMed source DOI: 10.1083/jcb.201109132 PMC Full Text View journal details
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