Abstract:
:The crystal structures of the GTP- and GDP-bound alpha subunits of heterotrimeric GTP-binding proteins were recently determined, and a conserved Thr residue in the G2 (linker 2) region of the alpha subunits, which corresponds to Thr182 in Gi2 alpha, was deduced to interact with the gamma-phosphate of GTP and Mg2+. To investigate biochemically the significance of the Thr residue, we produced a mutant Gi2 alpha, in which Thr182 was substituted for Ala (T182A), in Escherichia coli. The rate of guanosine 5'-(gamma-thio)tri-phosphate (GTP gamma S) binding to T182A was higher than that to the wild-type Gi2 alpha, especially with a high concentration (10 mM) of Mg2+. The rate of dissociation of bound GDP from T182A was also much faster than that from the wild-type with the high Mg2+ concentration. Moreover, T182A had much lower GTPase activity than the wild-type, like the gip mutant (R179C) of Gi2 alpha found in human endocrine tumors. The ability of T182A to interact with beta gamma subunits and membrane-bound receptors was the same as that of the wild-type alpha subunit. T182A could take on a GTP-bound active conformation, as judged from its sensitivity to tryptic digestion. These results indicated that Thr182 plays an important role not only in the Mg(2+)-sensitive GDP-GTP exchange reaction but also in the GTPase activity of Gi2 alpha. The T182A mutant of Gi2 alpha, characterized by the faster GDP release and the slower GTP hydrolysis, would be a novel mutant that retains the ability to interact with receptors and beta gamma subunits.
journal_name
J Biochemjournal_title
Journal of biochemistryauthors
Nishina H,Nimota K,Kukimoto I,Maehama T,Takahashi K,Hoshino S,Kanaho Y,Katada Tdoi
10.1093/jb/118.5.1083subject
Has Abstractpub_date
1995-11-01 00:00:00pages
1083-9issue
5eissn
0021-924Xissn
1756-2651journal_volume
118pub_type
杂志文章abstract::A comparative study of vertebrate ventricle tropomyosin has been carried out from the viewpoint of molecular evolution. The ventricles containing one-component tropomyosin were generally known, and in this paper those containing two components were also found in 8 species among mammals, reptiles, amphibia, and fish, b...
journal_title:Journal of biochemistry
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更新日期:1995-02-01 00:00:00
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更新日期:1978-08-01 00:00:00
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pub_type: 杂志文章
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更新日期:1982-12-01 00:00:00
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journal_title:Journal of biochemistry
pub_type: 杂志文章
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更新日期:1983-06-01 00:00:00
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pub_type: 杂志文章
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更新日期:1982-04-01 00:00:00
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pub_type: 杂志文章
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pub_type: 杂志文章
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pub_type: 杂志文章
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更新日期:1994-01-01 00:00:00
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更新日期:1985-04-01 00:00:00
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更新日期:1988-07-01 00:00:00
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更新日期:2002-10-01 00:00:00
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更新日期:2008-10-01 00:00:00
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更新日期:2019-06-14 00:00:00