Primary structure of a γ subunit of G protein, γ12, and its phosphorylation by protein kinase C

Rika Morishita, Hiroshi Nakayama, Toshiaki Isobe, Takahiko Matsuda, Yuichi Hashimoto, Toshiyuki Okano, Yoshitaka Fukada, Keiko Mizuno, Shigeo Ohno, Osamu Kozawa, Kanefusa Kato, Tomiko Asano

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Abstract

We have determined the primary structure of a novel γ subunit (γ12, previously designated γS1) of G protein purified from bovine spleen. The mature γ12 protein composed of 68 amino acids had acetylated serine at the N terminus and geranylgeranylated/carboxylmethylated cysteine at the C terminus. This was consistent with the C-terminal prenylation signal in the amino acid sequence, which was predicted from γ12 cDNA isolated from a bovine spleen cDNA library. Western blots with the specific antibody against γ12 showed that γ12 is present in all tissues examined. Among various γ subunits (γ1, γ2, γ3, Y7, and γ12), γ12 has a unique property to be phosphorylated by protein kinase C. The phosphorylated amino acid residue was Ser1 (or Ser2). The phosphorylated βγ12 associated with Goα more tightly than the unphosphorylated form. Exposure of Swiss 3T3 and aortic smooth muscle cells to phorbol 12-myristate 13-acetate and NaF induced phosphorylation of γ12. Stimulation of aortic smooth muscle cells with natural vasoactive agents such as angiotensin II and vasopressin also induced phosphorylation of γ12. The extent of phosphorylation of βγ12 in vitro was suppressed by a complex formation with Goα, which was relieved by the addition of guanosine 5′-O-(3-thiotriphosphate) or aluminum fluoride. These results strongly suggest that γ12 is phosphorylated by protein kinase C during activation of receptor(s) and G protein(s) in living cells.

Original languageEnglish
Pages (from-to)29469-29475
Number of pages7
JournalJournal of Biological Chemistry
Volume270
Issue number49
Publication statusPublished - 1995 Dec 8
Externally publishedYes

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Phosphorylation
GTP-Binding Proteins
Protein Kinase C
Cells
Amino Acids
Smooth Muscle Myocytes
Muscle
Spleen
Prenylation
Guanosine 5'-O-(3-Thiotriphosphate)
Vasopressins
Gene Library
Angiotensin II
Serine
Cysteine
Amino Acid Sequence
Acetates
Complementary DNA
Western Blotting
Chemical activation

ASJC Scopus subject areas

  • Biochemistry

Cite this

Morishita, R., Nakayama, H., Isobe, T., Matsuda, T., Hashimoto, Y., Okano, T., ... Asano, T. (1995). Primary structure of a γ subunit of G protein, γ12, and its phosphorylation by protein kinase C. Journal of Biological Chemistry, 270(49), 29469-29475.

Primary structure of a γ subunit of G protein, γ12, and its phosphorylation by protein kinase C. / Morishita, Rika; Nakayama, Hiroshi; Isobe, Toshiaki; Matsuda, Takahiko; Hashimoto, Yuichi; Okano, Toshiyuki; Fukada, Yoshitaka; Mizuno, Keiko; Ohno, Shigeo; Kozawa, Osamu; Kato, Kanefusa; Asano, Tomiko.

In: Journal of Biological Chemistry, Vol. 270, No. 49, 08.12.1995, p. 29469-29475.

Research output: Contribution to journalArticle

Morishita, R, Nakayama, H, Isobe, T, Matsuda, T, Hashimoto, Y, Okano, T, Fukada, Y, Mizuno, K, Ohno, S, Kozawa, O, Kato, K & Asano, T 1995, 'Primary structure of a γ subunit of G protein, γ12, and its phosphorylation by protein kinase C', Journal of Biological Chemistry, vol. 270, no. 49, pp. 29469-29475.
Morishita R, Nakayama H, Isobe T, Matsuda T, Hashimoto Y, Okano T et al. Primary structure of a γ subunit of G protein, γ12, and its phosphorylation by protein kinase C. Journal of Biological Chemistry. 1995 Dec 8;270(49):29469-29475.
Morishita, Rika ; Nakayama, Hiroshi ; Isobe, Toshiaki ; Matsuda, Takahiko ; Hashimoto, Yuichi ; Okano, Toshiyuki ; Fukada, Yoshitaka ; Mizuno, Keiko ; Ohno, Shigeo ; Kozawa, Osamu ; Kato, Kanefusa ; Asano, Tomiko. / Primary structure of a γ subunit of G protein, γ12, and its phosphorylation by protein kinase C. In: Journal of Biological Chemistry. 1995 ; Vol. 270, No. 49. pp. 29469-29475.
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AU - Fukada, Yoshitaka

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