End-Product Regulation and Kinetic Mechanism of Guanosine-Inosine Kinase from Escherichia coli

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概要

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• Escherichia coli guanosine-inosine kinase was over-produced, purified, and characterized. The native and subunit molecular weights were 85,000 and 45,000,respectively, indicating that the enzyme was a dimer. A pI of 6.0 was obtained by isoelectric focusing. In addition to ATP, it was found that deoxyadenosine 5'-triphosphate, UTP, and CTP could serve as phosphate donors. The phosphate acceptors were guanosine, inosine, deoxyguanosine and xanthosine, but not adenosine, cytidine, uridine, or deoxythymidine. Maximum activity was attained at an ATP/Mg^<2+> concentration ratio of 0.5. In the presence of pyrimidine nucleotides, enzyme activity was slightly increased, while it was markedly inhibited by GDP and GTP. Initial velocity and product inhibition studies support an ordered Bi Bi mechanism in which guanosine was the first substrate to bind and GMP was the last product to be released. Guanosine kinase may be a regulatory enzyme that has a role in modulating nucleotide levels.

• 社団法人日本農芸化学会の論文
• 2000-05-23

著者

• Kawasaki Haruhiko

  Department Of Applied Biochemistry Faculty Of Agriculture Osaka Prefecture University

• Kawasaki Haruhiko

  Department Of Applied Biochemistry Faculty Of Agriculture Osaka Prefecture Univeisity

• Kawasaki Haruhiko

  Department Of Applied Biochemistry Faculty Of Agriculture University Of Osaka Prefecture

• Kawasaki H

  Department Of Environmental Materials Science Tokyo Denki University

• Shimaoka Megumi

  Fermentation and Biotechnology Laboratories, Ajinomoto Co., Inc.

• Shimaoka M

  Fermentation And Biotechnology Laboratories Ajinomoto Co. Inc.

• KAWASAKI Hisashi

  Applied Microbiology Laboratory, Fermentation and Biotechnology Laboratories, Ajinomoto Co. Inc.

• USUDA Yoshihiro

  Applied Microbiology Laboratory, Fermentation and Biotechnology Laboratories, Ajinomoto Co. Inc.

• SHIMAOKA Megumi

  Applied Microbiology Laboratory, Fermentation and Biotechnology Laboratories, Ajinomoto Co. Inc.

• UTAGAWA Takashi

  Applied Microbiology Laboratory, Fermentation and Biotechnology Laboratories, Ajinomoto Co. Inc.

• Utagawa T

  Ajinomoto Co. Inc. Kawasaki‐shi Jpn

• Utagawa Takashi

  Applied Microbiology Laboratory Fermentation And Biotechnology Laboratories Ajinomoto Co. Inc.

• Usuda Yoshihiro

  Applied Microbiology Laboratory Fermentation And Biotechnology Laboratories Ajinomoto Co. Inc.

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