Docked secretory vesicles undergo Ca2+-activated exocytosis in a cell-free system.

The Journal of Biological Chemistry
Thomas F J Martin, Judith A Kowalchyk

Abstract

The Ca2+-activated fusion of secretory vesicles with the plasma membrane responsible for regulated neurotransmitter and hormone secretion has previously been studied in permeable neuroendocrine cells, where requirements for ATP and cytosolic proteins were identified. As reported here, Ca2+-activated fusion mechanisms are also preserved following cell homogenization. The release of norepinephrine (NE) and other vesicle constituents from a PC12 cell membrane fraction was activated by micromolar Ca2+ (EC50 approximately 3 microM) and exhibited a dependence upon MgATP and cytosol. Ca2+-dependent NE release was inhibited by botulinum neurotoxins and by CAPS (Ca2+-dependent activator protein for secretion) antibody implying that syntaxin, synaptobrevin, SNAP-25 (synaptosomal-associated protein of 25 kDa), and CAPS are required for regulated exocytosis in this system. The exocytosis-competent membrane fraction consisted of rapidly sedimenting dense core vesicles associated with plasma membrane fragments. Free vesicles did not release NE either in the absence or presence of plasma membranes, indicating that only docked vesicles were competent for exocytosis under the reconstitution conditions used. A cell-free system for Ca2+-activated...Continue Reading

References

Jan 1, 1989·Methods in Enzymology·T F Martin
Dec 1, 1985·The Journal of Cell Biology·J H Crabb, R C Jackson
Oct 1, 1985·Journal of Neurochemistry·J A Wagner
Nov 3, 1994·Nature·J E Rothman
Mar 25, 1993·Nature·T SöllnerJ E Rothman
Aug 1, 1995·Current Opinion in Structural Biology·L V Chernomordik, J Zimmerberg
Aug 23, 1996·The Journal of Biological Chemistry·A BanerjeeT F Martin
Aug 23, 1996·The Journal of Biological Chemistry·A BanerjeeT F Martin
Jan 1, 1996·Annual Review of Pharmacology and Toxicology·G J AugustineF E Schweizer
Jun 1, 1996·Current Opinion in Neurobiology·A W Henkel, W Almers
Nov 12, 1996·Proceedings of the National Academy of Sciences of the United States of America·G SchiavoJ E Rothman
Jun 1, 1997·Biologicals : Journal of the International Association of Biological Standardization·M Röllinghoff
May 1, 1994·Trends in Cell Biology·H NiemannR Jahn

❮ Previous
Next ❯

Citations

Jun 24, 2000·Biochimie·V A Klenchin, T F Martin
Aug 28, 1998·Current Opinion in Cell Biology·L J Robinson, T F Martin
Apr 15, 2008·Trends in Endocrinology and Metabolism : TEM·C Wiedemann, S Cockcroft
Mar 13, 2002·The EMBO Journal·William Wickner
Nov 20, 2002·Annals of the New York Academy of Sciences·James J Wassenberg, Thomas F J Martin
Aug 5, 2000·Applied and Environmental Microbiology·A G Zdanovsky, M V Zdanovskaia
Apr 1, 1999·Annual Review of Physiology·J AveryJ M Edwardson
Oct 27, 1999·Proceedings of the National Academy of Sciences of the United States of America·F ParlatiJ E Rothman
May 3, 2000·Proceedings of the National Academy of Sciences of the United States of America·M RupnikR Zorec
Jul 23, 2008·The Journal of Cell Biology·Declan J JamesThomas F J Martin
Dec 25, 2015·Molecular Biology of the Cell·Greg KabachinskiThomas F J Martin
Jan 29, 2000·The Journal of Cell Biology·J AveryR Jahn
Apr 4, 2006·Neurochemistry International·I O Trikash, L I Kolchinskaya
Aug 2, 2011·Cell Metabolism·Chuenchanok KhodthongThomas F J Martin
Sep 1, 2000·Seminars in Cell & Developmental Biology·R A Easom
Apr 23, 1999·Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences·H Kasai, N Takahashi
Oct 22, 2002·American Journal of Physiology. Cell Physiology·M KreftR Zorec
Jan 4, 2008·American Journal of Physiology. Cell Physiology·Haini LiaoDavid Castle
Feb 29, 2000·The Journal of Biological Chemistry·R R GeronaT F Martin
May 22, 2001·The Journal of Biological Chemistry·Y A ChenR H Scheller
Feb 24, 2001·Traffic·N R Cook, H W Davidson

❮ Previous
Next ❯

Related Concepts

Related Feeds

ASBMB Publications

The American Society for Biochemistry and Molecular Biology (ASBMB) includes the Journal of Biological Chemistry, Molecular & Cellular Proteomics, and the Journal of Lipid Research. Discover the latest research from ASBMB here.