Arg305 of streptomyces l-glutamate oxidase plays a crucial role for substrate recognition

Tomohiro Utsumi, Jiro Arima, Chika Sakaguchi, Takashi Tamura, Chiduko Sasaki, Hitoshi Kusakabe, Shigetoshi Sugio, Kenji Inagaki

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Recently, we have solved the crystal structure of l-glutamate oxidase (LGOX) from Streptomyces sp. X-119-6 (PDB code: 2E1M), the substrate specificity of which is strict toward l-glutamate. By a docking simulation using l-glutamate and structure of LGOX, we selected three residues, Arg305, His312, and Trp564 as candidates of the residues associating with recognition of l-glutamate. The activity of LGOX toward l-glutamate was significantly reduced by substitution of selected residues with Ala. However, the enzyme, Arg305 of which was substituted with Ala, exhibited catalytic activity toward various l-amino acids. To investigate the role of Arg305 in substrate specificity, we constructed Arg305 variants of LGOX. In all mutants, the substrate specificity of LGOX was markedly changed by the mutation. The results of kinetics and pH dependence on activity indicate that Arg305 of LGOX is associated with the interaction of enzyme and side chain of substrate.

Original languageEnglish
Pages (from-to)951-955
Number of pages5
JournalBiochemical and Biophysical Research Communications
Volume417
Issue number3
DOIs
Publication statusPublished - Jan 20 2012

Fingerprint

Streptomyces
Glutamic Acid
Oxidoreductases
Substrates
Substrate Specificity
Enzymes
Catalyst activity
Substitution reactions
Crystal structure
Amino Acids
Mutation
Kinetics

Keywords

  • L-Glutamate oxidase
  • Streptomyces
  • Substrate recognition
  • Substrate specificity

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Cell Biology
  • Molecular Biology

Cite this

Arg305 of streptomyces l-glutamate oxidase plays a crucial role for substrate recognition. / Utsumi, Tomohiro; Arima, Jiro; Sakaguchi, Chika; Tamura, Takashi; Sasaki, Chiduko; Kusakabe, Hitoshi; Sugio, Shigetoshi; Inagaki, Kenji.

In: Biochemical and Biophysical Research Communications, Vol. 417, No. 3, 20.01.2012, p. 951-955.

Research output: Contribution to journalArticle

Utsumi, Tomohiro ; Arima, Jiro ; Sakaguchi, Chika ; Tamura, Takashi ; Sasaki, Chiduko ; Kusakabe, Hitoshi ; Sugio, Shigetoshi ; Inagaki, Kenji. / Arg305 of streptomyces l-glutamate oxidase plays a crucial role for substrate recognition. In: Biochemical and Biophysical Research Communications. 2012 ; Vol. 417, No. 3. pp. 951-955.
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AU - Sasaki, Chiduko

AU - Kusakabe, Hitoshi

AU - Sugio, Shigetoshi

AU - Inagaki, Kenji

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