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PMID: 11879201 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Mechanism of the regulation of type IB phosphoinositide 3OH-kinase byG-protein betagamma subunits.

The Biochemical journal ·Vol. 362 ·No. Pt 3 ·2002-03-15 ·Pages 725-31

Krugmann S, Cooper MA, Williams DH, Hawkins PT, Stephens LR

Abstract

Type IB phosphoinositide 3OH-kinase (PI3K) is activated by G-protein betagamma subunits (Gbetagammas). The enzyme is soluble and largely cytosolic in vivo. Its substrate, PtdIns(4,5)P(2), and the Gbetagammas are localized at the plasma membrane. We have addressed the mechanism by which Gbetagammas regulate the PI3K using an in vitro approach. We used sedimentation assays and surface plasmon resonance to determine association of type IB PI3K with lipid monolayers and vesicles of varying compositions, some of which had Gbetagammas incorporated. Association and dissociation rate constants were determined. Our results indicated that in an assay situation in vitro the majority of PI3K will be associated with lipid vesicles, irrespective of the presence or absence of Gbetagammas. In line with this, a constitutively active membrane-targeted PI3K construct could still be activated substantially by Gbetagammas in vitro. We conclude that Gbetagammas activate type IB PI3K by a mechanism other than translocation to the plasma membrane.

MeSH Terms
Animals COS Cells Cell Line Chlorocebus aethiops Dimerization GTP-Binding Protein beta Subunits GTP-Binding Protein gamma Subunits Heterotrimeric GTP-Binding Proteins/metabolism Kinetics Phosphatidylinositol 3-Kinases/metabolism Protein Subunits Recombinant Proteins/metabolism Spodoptera Surface Plasmon Resonance Transfection
Chemicals
G-protein Beta gamma GTP-Binding Protein beta Subunits GTP-Binding Protein gamma Subunits Protein Subunits Recombinant Proteins Heterotrimeric GTP-Binding Proteins
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Krugmann Sonja
Signalling Programme, The Babraham Institute, Babraham, Cambridge CB2 4AT, U.K.
Cooper Matthew A
Williams Dudley H
Hawkins Phillip T
Stephens Len R
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Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
0264-6021
DOI
10.1042/0264-6021:3620725
Published
2002-03-15
Pages
725-31
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1222438
Subset
IM
External Links
PubMed source DOI: 10.1042/0264-6021:3620725 PMC Full Text View journal details
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