Use of specific antibodies to quantitate the guanine nucleotide-binding protein G(o) in brain

P. Gierschik, G. Milligan, M. Pines, P. Goldsmith, J. Codina, W. Klee, Allen M. Spiegel

Research output: Contribution to journalArticle

113 Citations (Scopus)

Abstract

We immunized rabbits with purified guanine nucleotide-binding proteins (G proteins) from bovine brain and obtained an antiserum, RV3, that reacts specifically with the α subunit (39 kDa) of a G protein of unknown function, termed G(o), as well as with the β subunit (35 kDa) common to all G proteins. RV3 showed no crossreactivity with the α subunits of the stimulatory (G(s) or inhibitory (G(i)) G proteins associated with adenylate cyclase, nor with that of the rod outer segment G protein, transducin. Immunoblots with crude and affinity-purified antiserum showed that RV3 specifically recognizes the G(o) α subunit and the β subunit in crude brain membranes. Using RV3, we found approximately equal amounts of G(o) in brain membranes from frog, chicken, rat, cow, and man. Quantitative immunoblotting gave G(o) α subunit/β subunit ratios ~ 1 in cerebral cortex, raising the possibility that free G(o) α subunit (unassociated with β subunit) may exist in brain. The concentration of G(o) α subunit in cerebral cortex is about 5 times that of G(i) α subunit. The results show that G(o) is an immunochemically distinct, highly conserved protein distributed throughout the brain, with particularly high concentrations in forebrain.

Original languageEnglish (US)
Pages (from-to)2258-2262
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume83
Issue number7
StatePublished - 1986
Externally publishedYes

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Guanine Nucleotides
GTP-Binding Proteins
Carrier Proteins
Antibodies
Brain
Cerebral Cortex
Immune Sera
Transducin
Rod Cell Outer Segment
Membranes
Prosencephalon
Immunoblotting
Adenylyl Cyclases
Anura
Chickens
Rabbits
Proteins

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Use of specific antibodies to quantitate the guanine nucleotide-binding protein G(o) in brain. / Gierschik, P.; Milligan, G.; Pines, M.; Goldsmith, P.; Codina, J.; Klee, W.; Spiegel, Allen M.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 83, No. 7, 1986, p. 2258-2262.

Research output: Contribution to journalArticle

Gierschik, P. ; Milligan, G. ; Pines, M. ; Goldsmith, P. ; Codina, J. ; Klee, W. ; Spiegel, Allen M. / Use of specific antibodies to quantitate the guanine nucleotide-binding protein G(o) in brain. In: Proceedings of the National Academy of Sciences of the United States of America. 1986 ; Vol. 83, No. 7. pp. 2258-2262.
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