Purification and partial characterization of 3-hydroxyisobutyryl-coenzyme A hydrolase of rat liver

Yoshiharu Shimomura, Taro Murakami, Noriaki Fujitsuka, Naoya Nakai, Yuzo Sato, Satoru Sugiyama, Noriko Shimomura, Jamie Irwin, John W. Hawes, Robert Harris

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Abstract

An unusual feature of valine catabolism is a reaction in which an intermediate of its catabolic pathway, (S)-3-hydroxyisobutyryl-CoA, is hydrolyzed to give the free acid and CoA-SH. The enzyme responsible for this reaction, 3-hydroxyisobutyryl-CoA hydrolase (EC 3.1.2.4), was purified 7200- fold from rat liver in this study. The purified enzyme consists of a single polypeptide with an M(r) of 36,000 in the native and denatured forms. The hydrolase is highly specific for (S)-3-hydroxyisobutyryl-CoA and 3- hydroxypropionyl-CoA (K(m), 6 and 25 μM, respectively) with optimal activity around pH 8. The turnover rate of the enzyme for (S)-3-hydroxyisobutyryl-CoA is 270 s-1, which is high relative to other enzymes of the valine pathway. Likewise, activity of the enzyme expressed on a wet weight basis is also very high in the major tissues of the rat. These findings suggest that rapid destruction of (S)-3-hydroxyisobutyryl-CoA produced during valine catabolism is physiologically important. We propose that the need for a mechanism to protect cells against the toxic effects of methacrylyl-CoA, which is maintained in equilibrium with (S)-3-hydroxyisobutyryl-CoA by crotonase, explains why valine catabolism involves this enzyme and why its tissue activity is so high.

Original languageEnglish
Pages (from-to)14248-14253
Number of pages6
JournalJournal of Biological Chemistry
Volume269
Issue number19
StatePublished - May 13 1994

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3-hydroxyisobutyryl-CoA hydrolase
Coenzyme A
Liver
Purification
Rats
Valine
Enzymes
Enoyl-CoA Hydratase
Tissue
Poisons
Hydrolases

ASJC Scopus subject areas

  • Biochemistry

Cite this

Shimomura, Y., Murakami, T., Fujitsuka, N., Nakai, N., Sato, Y., Sugiyama, S., ... Harris, R. (1994). Purification and partial characterization of 3-hydroxyisobutyryl-coenzyme A hydrolase of rat liver. Journal of Biological Chemistry, 269(19), 14248-14253.

Purification and partial characterization of 3-hydroxyisobutyryl-coenzyme A hydrolase of rat liver. / Shimomura, Yoshiharu; Murakami, Taro; Fujitsuka, Noriaki; Nakai, Naoya; Sato, Yuzo; Sugiyama, Satoru; Shimomura, Noriko; Irwin, Jamie; Hawes, John W.; Harris, Robert.

In: Journal of Biological Chemistry, Vol. 269, No. 19, 13.05.1994, p. 14248-14253.

Research output: Contribution to journalArticle

Shimomura, Y, Murakami, T, Fujitsuka, N, Nakai, N, Sato, Y, Sugiyama, S, Shimomura, N, Irwin, J, Hawes, JW & Harris, R 1994, 'Purification and partial characterization of 3-hydroxyisobutyryl-coenzyme A hydrolase of rat liver', Journal of Biological Chemistry, vol. 269, no. 19, pp. 14248-14253.
Shimomura Y, Murakami T, Fujitsuka N, Nakai N, Sato Y, Sugiyama S et al. Purification and partial characterization of 3-hydroxyisobutyryl-coenzyme A hydrolase of rat liver. Journal of Biological Chemistry. 1994 May 13;269(19):14248-14253.
Shimomura, Yoshiharu ; Murakami, Taro ; Fujitsuka, Noriaki ; Nakai, Naoya ; Sato, Yuzo ; Sugiyama, Satoru ; Shimomura, Noriko ; Irwin, Jamie ; Hawes, John W. ; Harris, Robert. / Purification and partial characterization of 3-hydroxyisobutyryl-coenzyme A hydrolase of rat liver. In: Journal of Biological Chemistry. 1994 ; Vol. 269, No. 19. pp. 14248-14253.
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