Purification of heterotrimeric GTP-binding proteins from brain: Identification of a novel form of Go

Paul Goldsmith, Peter S. Backlund, Kevin Rossiter, Anthony Carter, Graeme Milligan, Allen M. Spiegel, Allen Spiegel

Research output: Contribution to journalArticle

107 Citations (Scopus)

Abstract

Using high-resolution Mono-Q anion-exchange chromatography, we purified four distinct GTP-binding proteins from bovine brain. Each consists of α and associated β/γ subunits, and each is a substrate for pertussis toxin catalyzed ADP-ribosylation. We defined the relationship between the a subunits of the purified proteins and cloned cDNAs encoding putative α subunits (1) by performing immunoblots with peptide antisera with defined specificity and (2) by comparing the migration on two-dimensional gel electrophoresis of the purified proteins, and of the in vitro translated products of cDNAs encoding α subunits. Purified G proteins with α subunits of 39, 41, and 40 kDa (G39, G41, and G40 in order of abundance) correspond to the products of Go, Gil, and Gi2 cDNAs. We purified a novel G protein with an a subunit slightly above 39 kDa (G39*). G39* is less abundant than G39, elutes earlier than G39 on Mono-Q chromatography, and has a more basic pI (6.0 vs 5.6) than G39. G39 and G39*, however, are indistinguishable on immunoblots with a large number of specific antisera. The data suggest that G39* may represent a novel form of Go, differing in posttranslational modification rather than primary sequence.

Original languageEnglish (US)
Pages (from-to)7085-7090
Number of pages6
JournalBiochemistry
Volume27
Issue number18
StatePublished - 1988
Externally publishedYes

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Heterotrimeric GTP-Binding Proteins
GTP-Binding Proteins
Purification
Brain
Complementary DNA
Protein Subunits
Chromatography
Immune Sera
Pertussis Toxin
Electrophoresis, Gel, Two-Dimensional
Post Translational Protein Processing
Electrophoresis
Adenosine Diphosphate
Anions
Proteins
Gels
Peptides
Substrates
Mono Q

ASJC Scopus subject areas

  • Biochemistry

Cite this

Goldsmith, P., Backlund, P. S., Rossiter, K., Carter, A., Milligan, G., Spiegel, A. M., & Spiegel, A. (1988). Purification of heterotrimeric GTP-binding proteins from brain: Identification of a novel form of Go. Biochemistry, 27(18), 7085-7090.

Purification of heterotrimeric GTP-binding proteins from brain : Identification of a novel form of Go. / Goldsmith, Paul; Backlund, Peter S.; Rossiter, Kevin; Carter, Anthony; Milligan, Graeme; Spiegel, Allen M.; Spiegel, Allen.

In: Biochemistry, Vol. 27, No. 18, 1988, p. 7085-7090.

Research output: Contribution to journalArticle

Goldsmith, P, Backlund, PS, Rossiter, K, Carter, A, Milligan, G, Spiegel, AM & Spiegel, A 1988, 'Purification of heterotrimeric GTP-binding proteins from brain: Identification of a novel form of Go', Biochemistry, vol. 27, no. 18, pp. 7085-7090.
Goldsmith P, Backlund PS, Rossiter K, Carter A, Milligan G, Spiegel AM et al. Purification of heterotrimeric GTP-binding proteins from brain: Identification of a novel form of Go. Biochemistry. 1988;27(18):7085-7090.
Goldsmith, Paul ; Backlund, Peter S. ; Rossiter, Kevin ; Carter, Anthony ; Milligan, Graeme ; Spiegel, Allen M. ; Spiegel, Allen. / Purification of heterotrimeric GTP-binding proteins from brain : Identification of a novel form of Go. In: Biochemistry. 1988 ; Vol. 27, No. 18. pp. 7085-7090.
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