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PMID: 19460754 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

CAPS activity in priming vesicle exocytosis requires CK2 phosphorylation.

The Journal of biological chemistry ·Vol. 284 ·No. 28 ·2009-07-10 ·Pages 18707-14

Nojiri M, Loyet KM, Klenchin VA, Kabachinski G, Martin TF

Abstract

CAPS (Ca(2+)-dependent activator protein for secretion) functions in priming Ca(2+)-dependent vesicle exocytosis, but the regulation of CAPS activity has not been characterized. Here we show that phosphorylation by protein kinase CK2 is required for CAPS activity. Dephosphorylation eliminated CAPS activity in reconstituting Ca(2+)-dependent vesicle exocytosis in permeable and intact PC12 cells. Ser-5, -6, and -7 and Ser-1281 were identified by mass spectrometry as the major phosphorylation sites in the 1289 residue protein. Ser-5, -6, and -7 but not Ser-1281 to Ala substitutions abolished CAPS activity. Protein kinase CK2 phosphorylated CAPS in vitro at these sites and restored the activity of dephosphorylated CAPS. CK2 is the likely in vivo CAPS protein kinase based on inhibition of phosphorylation by tetrabromo-2-benzotriazole in PC12 cells and by the identity of in vivo and in vitro phosphorylation sites. CAPS phosphorylation by CK2 was constitutive, but the elevation of Ca(2+) in synaptosomes increased CAPS Ser-5 and -6 dephosphorylation, which terminates CAPS activity. These results identify a functionally important N-terminal phosphorylation site that regulates CAPS activity in priming vesicle exocytosis.

MeSH Terms
Amino Acid Sequence Animals COS Cells Calcium/chemistry Calcium-Binding Proteins/metabolism,physiology Casein Kinase II/chemistry,metabolism Chlorocebus aethiops Exocytosis Humans Mice Molecular Sequence Data PC12 Cells Phosphorylation Rats Sequence Homology, Amino Acid Serine/chemistry Triazoles/pharmacology
Chemicals
Cadps protein, rat Calcium-Binding Proteins Triazoles Serine benzotriazole Casein Kinase II Calcium
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Nojiri Mari
Department of Biochemistry, University of Wisconsin, Madison, Wisconsin 53706, USA.
Loyet Kelly M
Klenchin Vadim A
Kabachinski Gregory
Martin Thomas F J
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Article Info
Journal
The Journal of biological chemistry
Abbr.
J Biol Chem
ISSN
0021-9258
DOI
10.1074/jbc.M109.017483
Published
2009-07-10
Epub
2009-00-21
Pages
18707-14
Language
English
Region
United States
NLM ID
2985121R
PMCID
PMC2707249
Subset
IM
Grants
NIDDK NIH HHS · DK25861 · United States
NIDDK NIH HHS · R01 DK040428 · United States
NIDDK NIH HHS · R01 DK025861 · United States
NIDDK NIH HHS · DK40428 · United States
NIDDK NIH HHS · R37 DK025861 · United States
External Links
PubMed source DOI: 10.1074/jbc.M109.017483 PMC Full Text View journal details
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