Biochemical characterization of two glutamate dehydrogenases with different cofactor specificities from a hyperthermophilic archaeon Pyrobaculum calidifontis

Taisuke Wakamatsu, Chisato Higashi, Taketo Ohmori, Katsumi Doi, Toshihisa Ohshima

研究成果: ジャーナルへの寄稿記事

2 引用 (Scopus)

抄録

Two putative glutamate dehydrogenase (GDH) genes (pcal_1031 and pcal_1606) were found in a sulfur-dependent hyperthermophilic archaeon, Pyrobaculum calidifontis. The two genes were then expressed in Escherichia coli, and both of the recombinant gene products showed GDH activity. The two enzymes were then purified to homogeneity and characterized in detail. Although both purified GDHs had a hexameric structure and neither exhibited allosteric regulation, they showed different coenzyme specificities: one was specific for NAD+, the other for NADP+ and different heat activation mechanisms. In addition, there was little difference in the kinetic constants, optimal temperature, thermal stability, optimal pH and pH stability between the two enzymes. The overall sequence identity between the two proteins was very high (81 %), but was not high in the region recognizing the 2′ position of the adenine ribose moiety, which is responsible for coenzyme specificity. This is the first report on the identification of two GDHs with different coenzyme specificities from a single hyperthermophilic archaeon and the definition of their basic in vitro properties.

元の言語英語
ページ(範囲)379-389
ページ数11
ジャーナルExtremophiles
17
発行部数3
DOI
出版物ステータス出版済み - 3 19 2013

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Pyrobaculum
Glutamate Dehydrogenase
Coenzymes
Archaea
Hot Temperature
Allosteric Regulation
Genes
Ribose
Adenine
Enzymes
NADP
Sulfur
NAD
Escherichia coli
Temperature
Proteins

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Molecular Medicine

これを引用

Biochemical characterization of two glutamate dehydrogenases with different cofactor specificities from a hyperthermophilic archaeon Pyrobaculum calidifontis. / Wakamatsu, Taisuke; Higashi, Chisato; Ohmori, Taketo; Doi, Katsumi; Ohshima, Toshihisa.

:: Extremophiles, 巻 17, 番号 3, 19.03.2013, p. 379-389.

研究成果: ジャーナルへの寄稿記事

Wakamatsu, Taisuke ; Higashi, Chisato ; Ohmori, Taketo ; Doi, Katsumi ; Ohshima, Toshihisa. / Biochemical characterization of two glutamate dehydrogenases with different cofactor specificities from a hyperthermophilic archaeon Pyrobaculum calidifontis. :: Extremophiles. 2013 ; 巻 17, 番号 3. pp. 379-389.
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abstract = "Two putative glutamate dehydrogenase (GDH) genes (pcal_1031 and pcal_1606) were found in a sulfur-dependent hyperthermophilic archaeon, Pyrobaculum calidifontis. The two genes were then expressed in Escherichia coli, and both of the recombinant gene products showed GDH activity. The two enzymes were then purified to homogeneity and characterized in detail. Although both purified GDHs had a hexameric structure and neither exhibited allosteric regulation, they showed different coenzyme specificities: one was specific for NAD+, the other for NADP+ and different heat activation mechanisms. In addition, there was little difference in the kinetic constants, optimal temperature, thermal stability, optimal pH and pH stability between the two enzymes. The overall sequence identity between the two proteins was very high (81 {\%}), but was not high in the region recognizing the 2′ position of the adenine ribose moiety, which is responsible for coenzyme specificity. This is the first report on the identification of two GDHs with different coenzyme specificities from a single hyperthermophilic archaeon and the definition of their basic in vitro properties.",
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