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PMID: 12821674 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Calmodulin is a phospholipase C-beta interacting protein.

The Journal of biological chemistry ·Vol. 278 ·No. 36 ·2003-09-05 ·Pages 33708-13

McCullar JS, Larsen SA, Millimaki RA, Filtz TM

Abstract

Phospholipase C-beta 3 (PLC beta 3) is an important effector enzyme in G protein-coupled signaling pathways. Activation of PLC beta 3 by G alpha and G beta gamma subunits has been fairly well characterized, but little is known about other protein interactions that may also regulate PLC beta 3 function. A yeast two-hybrid screen of a mouse brain cDNA library with the amino terminus of PLC beta 3 has yielded potential PLC beta 3 interacting proteins including calmodulin (CaM). Physical interaction between CaM and PLC beta 3 is supported by a positive secondary screen in yeast and the identification of a CaM binding site in the amino terminus of PLC beta 3. Co-precipitation of in vitro translated and transcribed amino- and carboxyl-terminal PLC beta 3 revealed CaM binding at a putative amino-terminal binding site. Direct physical interaction of PLC beta 3 and PLC beta 1 isoforms with CaM is supported by pull-down of both isoenzymes with CaM-Sepharose beads from 1321N1 cell lysates. CaM inhibitors reduced M1-muscarinic receptor stimulation of inositol phospholipid hydrolysis in 1321N1 astrocytoma cells consistent with a physiologic role for CaM in modulation of PLC beta activity. There was no effect of CaM kinase II inhibitors, KN-93 and KN-62, on M1-muscarinic receptor stimulation of inositol phosphate hydrolysis, consistent with a direct interaction between PLC beta isoforms and CaM.

MeSH Terms
Amino Acid Sequence Animals Binding Sites Calmodulin/chemistry,metabolism,physiology Carbachol/chemistry DNA, Complementary/metabolism Gene Library Humans Hydrolysis Inositol Phosphates/metabolism Isoenzymes/metabolism Mice Molecular Sequence Data Phospholipase C beta Precipitin Tests Protein Binding Protein Biosynthesis Protein Isoforms Protein Structure, Tertiary Sepharose/chemistry Sequence Homology, Amino Acid Signal Transduction Subcellular Fractions/metabolism Transcription, Genetic Tumor Cells, Cultured Two-Hybrid System Techniques Type C Phospholipases/metabolism beta-Galactosidase/metabolism
Chemicals
Calmodulin DNA, Complementary Inositol Phosphates Isoenzymes Protein Isoforms Carbachol Sepharose Type C Phospholipases PLCB1 protein, human PLCB3 protein, human Phospholipase C beta Plcb1 protein, mouse Plcb3 protein, mouse beta-Galactosidase
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
McCullar Jennifer S
Department of Pharmaceutical Sciences, College of Pharmacy, and the Molecular and Cellular Biology Program, Oregon State University, Corvallis, Oregon 97331, USA.
Larsen Shana A
Millimaki Ryan A
Filtz Theresa M
Article Info
Journal
The Journal of biological chemistry
Abbr.
J Biol Chem
ISSN
0021-9258
Published
2003-09-05
Epub
2003-00-23
Pages
33708-13
Language
English
Region
United States
NLM ID
2985121R
Subset
IM
Grants
NIGMS NIH HHS · R01 GM061244 · United States
NIGMS NIH HHS · GM61244 · United States
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