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Membrane attachment of recombinant G-protein α-subunits in excess of βγ subunits in a eukaryotic expression system

Authors
Journal
Biochemical and Biophysical Research Communications
0006-291X
Publisher
Elsevier
Publication Date
Volume
164
Issue
1
Identifiers
DOI: 10.1016/0006-291x(89)91680-x

Abstract

Abstract Recombinant cDNAs encoding the α-subunits of G i1, G i2, G i3, G O and G S were transfected into COS cells with the pCD-PS mammalian expression vector. Expression of each G α was verified using subtype-specific peptide antisera on immunoblots. Quantitative immunoblotting of α and β subunits indicated: i) that there was no change in expression of endogenous β subunits, and ii) overexpression of α subunits could achieve a ratio of α:β > 25:1. Despite the excess of α over β, the G α subunits were found predominantly in the membrane fraction. The results demonstrate that G α subunits can attach to the membrane independently of βγ subunits.

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