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PMID: 2165390 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Isolation and characterization of a gamma-type phosphoinositide-specific phospholipase C (PLC-gamma 2).

The Biochemical journal ·Vol. 269 ·No. 1 ·1990-07-01 ·Pages 13-8

Homma Y, Emori Y, Shibasaki F, Suzuki K, Takenawa T

Abstract

A novel bovine spleen phosphoinositide-specific phospholipase C (PLC) has been identified with respect to immunoreactivity with four independent antibodies against each of the PLC isoenzymes, and purified to near homogeneity by sequential column chromatography. Spleen contains three of the isoenzymes: two different gamma-types [gamma 1 and gamma 2, originally named as PLC-gamma [Rhee, Suh, Ryu & Lee (1989) Science 244, 546-550] and PLC-IV [Emori, Homma, Sorimachi, Kawasaki, Nakanishi, Suzuki & Takenawa (1989) J. Biol. Chem. 264, 21885-21890] respectively] and delta-type of the enzyme, but PLC-gamma 1 is separated from the PLC-gamma 2 pool by the first DEAE-cellulose column chromatography. Subsequently, PLC-delta is dissociated on the third heparin-Sepharose column chromatography. The purified enzyme has a molecular mass of 145 kDa on SDS/polyacrylamide-gel electrophoresis and a specific activity of 12.8 mumol/min per mg with phosphatidylinositol 4,5-bisphosphate as substrate. This enzyme activity is dependent on Ca2+ for hydrolysis of all these phosphoinositides. None of the other phospholipids examined could be its substrate at any concentration of Ca2+. The optimal pH of the enzyme is slightly acidic (pH 5.0-6.5).

MeSH Terms
Animals Calcium/pharmacology Cattle Chromatography Electrophoresis, Polyacrylamide Gel Hydrogen-Ion Concentration Immunoblotting Isoenzymes/isolation & purification,metabolism Kinetics Molecular Weight Phosphatidylinositol 4,5-Diphosphate Phosphatidylinositols/metabolism Spleen/enzymology Substrate Specificity Type C Phospholipases/isolation & purification,metabolism
Chemicals
Isoenzymes Phosphatidylinositol 4,5-Diphosphate Phosphatidylinositols Type C Phospholipases Calcium
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Homma Y
Department of Pharmacology, Tokyo Metropolitan Institute of Gerontology, Japan.
Emori Y
Shibasaki F
Suzuki K
Takenawa T
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21 references, click to expand
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Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
0264-6021
DOI
10.1042/bj2690013
Published
1990-07-01
Pages
13-8
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1131523
Subset
IM
External Links
PubMed source DOI: 10.1042/bj2690013 PMC Full Text View journal details
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