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

William F. Simonds, Regina M. Collins, Allen M. Spiegel, Mark R. Brann

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

Recombinant cDNAs encoding the α-subunits of Gi1, Gi2, Gi3, GO and GS were transfected into COS cells with the pCD-PS mammalian expression vector. Expression of each Gα 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 Gα subunits were found predominantly in the membrane fraction. The results demonstrate that Gα subunits can attach to the membrane independently of βγ subunits.

Original languageEnglish (US)
Pages (from-to)46-53
Number of pages8
JournalBiochemical and Biophysical Research Communications
Volume164
Issue number1
DOIs
StatePublished - Oct 16 1989
Externally publishedYes

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Protein Subunits
GTP-Binding Proteins
Recombinant Proteins
Membranes
COS Cells
Immunoblotting
Immune Sera
Complementary DNA
Peptides

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Membrane attachment of recombinant G-protein α-subunits in excess of βγ subunits in a eukaryotic expression system. / Simonds, William F.; Collins, Regina M.; Spiegel, Allen M.; Brann, Mark R.

In: Biochemical and Biophysical Research Communications, Vol. 164, No. 1, 16.10.1989, p. 46-53.

Research output: Contribution to journalArticle

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