Isolation of two proteins with high affinity for guanine nucleotides from membranes of bovine brain

P. C. Sternweis, J. D. Robishaw

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Abstract

Membranes from bovine brain bind relatively large quantities of guanosine 5'-(3-O-thio)triphosphate (GTPγS) with high affinity. The two proteins responsible for most of this activity were purified; they account for 1.5% of the membrane protein. The two proteins contain α subunits of either 39,000 and 41,000 Da, β subunits of 36,000 or 35,000 Da, and a potential γ subunit (11,000 Da). These structures are the same as a family of proteins that includes transducin and the regulatory proteins, G(S) and G(I), of adenylate cyclase. The 41,000- and 39,000-Da polypeptides can be ADP-ribosylated with islet-activating protein from Bordetella pertussis, bind guanine nucleotides specifically, and migrate through polyacrylamide gels with rates similar to the α subunits of G(I) and transducin, respectively. The 36,000- and 35,000-Da polypeptides are similar to the β subunits of G(I) and G(S). The γ subunit is found whenever β subunits are present. The 41,000- and 39,000-Da polypeptides (with β and γ) are designated, respectively, G(I) and G(O) from brain. The α subunit of G(O) was isolated without the use of ligands known to dissociate other G proteins. G(O)α binds GTPγS reversible in the absence of Mg2+ and is relatively stable in cholate. This isolated α subunit should be of great utility in elucidating the mechanism of action of this family of GTP-binding proteins.

Original languageEnglish (US)
Pages (from-to)13806-13813
Number of pages8
JournalJournal of Biological Chemistry
Volume259
Issue number22
StatePublished - 1984

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Guanine Nucleotides
Transducin
Brain
Membranes
GTP-Binding Proteins
Peptides
Cholates
Guanosine 5'-O-(3-Thiotriphosphate)
Bordetella pertussis
Proteins
Protein S
Pertussis Toxin
Protein Subunits
Adenylyl Cyclases
Adenosine Diphosphate
Membrane Proteins
Ligands

ASJC Scopus subject areas

  • Biochemistry

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Isolation of two proteins with high affinity for guanine nucleotides from membranes of bovine brain. / Sternweis, P. C.; Robishaw, J. D.

In: Journal of Biological Chemistry, Vol. 259, No. 22, 1984, p. 13806-13813.

Research output: Contribution to journalArticle

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