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PMID: 7622570 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

A phosphatidylinositol 4,5-bisphosphate-sensitive casein kinase I alpha associates with synaptic vesicles and phosphorylates a subset of vesicle proteins.

The Journal of cell biology ·Vol. 130 ·No. 3 ·1995-08-00 ·Pages 711-24

Gross SD, Hoffman DP, Fisette PL, Baas P, Anderson RA

Abstract

In interphase cells, alpha-casein kinase I (alpha-CKI) is found associated with cytosolic vesicular structures, the centrosome, and within the nucleus. To identify the specific vesicular structures with which alpha-CKI is associated, established cell lines and primary rat neurons were immunofluorescently labeled with an antibody raised to alpha-CKI. In nonneuronal cells, alpha-CKI colocalizes with vesicular structures which align with microtubules and are partially coincident with both Golgi and endoplasmic reticulum markers. In neurons, alpha-CKI colocalizes with synaptic vesicle markers. When synaptic vesicles were purified from rat brain, they were highly enriched in a CKI, based on activity and immunoreactivity. The synaptic vesicle-associated CKI is an extrinsic kinase and was eluted from synaptic vesicles and purified. This purified CKI has properties most similar to alpha-CKI. When the activities of casein kinase I or II were specifically inhibited on isolated synaptic vesicles, CKI was shown to phosphorylate a specific subset of vesicle proteins, one of which was identified as the synaptic vesicle-specific protein SV2. As with alpha-CKI, the synaptic vesicle CKI is inhibited by phosphatidylinositol 4,5-bisphosphate (PIP2). However, synthesis of PIP2 was detected only in plasma membrane-containing fractions. Therefore, PIP2 may spatially regulate CKI. Since PIP2 synthesis is required for secretion, this inhibition of CKI may be important for the regulation of secretion.

MeSH Terms
Animals Biological Transport Blotting, Western Brain/enzymology CHO Cells Casein Kinases Cell Fractionation Cell Membrane/metabolism Cricetinae Fluorescent Antibody Technique Kidney/cytology Membrane Glycoproteins/metabolism Microscopy, Electron Nerve Tissue Proteins/metabolism Neurons/physiology,ultrastructure Neurotransmitter Agents/metabolism PC12 Cells Phosphatidylinositol 4,5-Diphosphate Phosphatidylinositol Phosphates/pharmacology Phosphorylation Precipitin Tests Protein Binding Protein Kinases/drug effects,isolation & purification,metabolism Rats Swine Synaptic Transmission/physiology Synaptic Vesicles/metabolism
Chemicals
Membrane Glycoproteins Nerve Tissue Proteins Neurotransmitter Agents Phosphatidylinositol 4,5-Diphosphate Phosphatidylinositol Phosphates Sv2a protein, rat Protein Kinases Casein Kinases
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Gross S D
Department of Pharmacology, University of Wisconsin Medical School, Madison 53706, USA.
Hoffman D P
Fisette P L
Baas P
Anderson R A
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.130.3.711
Published
1995-08-00
Pages
711-24
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2120523
Subset
IM
Grants
NIGMS NIH HHS · GM38906 · United States
NIGMS NIH HHS · GM51968 · United States
External Links
PubMed source DOI: 10.1083/jcb.130.3.711 PMC Full Text View journal details
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