The primary structure of the α subunit of a starfish guanosine-nucleotide-binding regulatory protein involved in 1-methyladenine-induced oocyte maturation

K. Chiba, H. Tadenuma, Midori Matsumoto, K. Takahashi, T. Katada, M. Hoshi

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Abstract

Starfish-oocyte maturation induced by 1-methyladenine (MeAde) was inhibited by microinjection of pertussis toxin (PTX). The inhibition appeared to result from PTX-catalyzed ADP-ribosylation of a 39-kDa guanosine-nucleotide-binding regulatory protein (G protein) in the oocyte. These results strongly support the hypothesis that the MeAde-induced signals operate via a membrane receptor and are carried by the PTX-sensitive G protein. When PTX-injected oocytes were treated with dithiothreitol, 85% of them reinitiated meiosis, suggesting that dithiothreitol did not act on the MeAde receptor. We constructed a cDNA library from the immature ovary of starfish, Asterina pectinifera, and screened it with the cDNA of the α subunit of an inhibitory rat G protein (G(i-2)). A positive cDNA clone contained an open reading frame of 1062 bases which had 74% identity with the rat G(i-2) cDNA. The deduced amino acid sequence was 85% and 89% identical to rat G(i-2) and rat G(i-1), respectively. The α subunit of the G protein purified from cortices of starfish oocytes was digested by trypsin and the resulting four peptides were microsequenced. Comparison of these amino acid sequences with the predicted one indicated that the isolated cDNA clone encoded the α subunit of the PTX-sensitive G protein in oocytes. The C-terminal sequence, KNNLKDCGLF, was identical to that of G(i), suggesting that the cysteine residue is the site of ADP-ribosylation.

Original languageEnglish
Pages (from-to)833-838
Number of pages6
JournalEuropean Journal of Biochemistry
Volume207
Issue number3
DOIs
Publication statusPublished - 1992
Externally publishedYes

Fingerprint

Starfish
Guanosine
Oocytes
Pertussis Toxin
Carrier Proteins
Nucleotides
Rats
Complementary DNA
Dithiothreitol
Proteins
Adenosine Diphosphate
Amino Acid Sequence
Asterina
Clone Cells
Amino Acids
Meiosis
Microinjections
Gene Library
Trypsin
Open Reading Frames

ASJC Scopus subject areas

  • Biochemistry

Cite this

The primary structure of the α subunit of a starfish guanosine-nucleotide-binding regulatory protein involved in 1-methyladenine-induced oocyte maturation. / Chiba, K.; Tadenuma, H.; Matsumoto, Midori; Takahashi, K.; Katada, T.; Hoshi, M.

In: European Journal of Biochemistry, Vol. 207, No. 3, 1992, p. 833-838.

Research output: Contribution to journalArticle

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