Mutational Study on αGln90 of Fe-Type Nitrile Hydratase from Rhodococcus sp. N771
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概要
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Nitrile hydratase (NHase) from Rhodococcus sp. N771 is a non-heme iron enzyme having post-translationally modified cysteine ligands, αCys112-SO2H and αCys114-SOH. We replaced αGln90, which is conserved in all known NHases and involved in the hydrogen-bond network around the catalytic center, with glutamic acid or asparagine. The kcat of αQ90E and αQ90N mutants decreased to 24% and 5% that of wild type respectively, but the effect of mutations on Km was not very significant. In both mutants, the αCys114-SOH modification appeared to be responsible for the catalysis as in native NHase. We crystallized the nitrosylated αQ90N mutant and determined its structure at a resolution of 1.43 Å. The structure was basically identical to that of native nitrosylated NHase except for the mutated site and its vicinity. The structural difference between native and αQ90N mutant NHases suggested the importance of the hydrogen bond networks between αGln90 and the iron center for the catalytic activity.
- 社団法人 日本農芸化学会の論文
- 2006-04-23
著者
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DOHMAE Naoshi
Biomolecular Characterization Team, RIKEN
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Dohmae Naoshi
Biomolecular Characterization Team Riken
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Maeda Mizuo
Maeda Bioengineering Laboratory Riken
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Yohda Masafumi
Department Of Biotechnology And Life Science Graduate School Of Engineering Tokyo University Of Agri
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Kamiya Nobuo
Protein Crystallography Methodology Team Advanced Protein Crystallography Research Group Riken Harim
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Dohmae Naoshi
Biomolecular Characterization Division Riken
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Odaka Masafumi
Maeda Bioengineering Laboratory Riken:department Of Biotechnology And Life Science Graduate School O
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TAKARADA Hiromi
United Graduate School of Agricultural Science, Tokyo University of Agriculture and Technology
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KAWANO Yoshiaki
Protein Crystallography Methodology Team, Advanced Protein Crystallography Research Group, RIKEN Har
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HASHIMOTO Kouichi
Protein Crystallography Methodology Team, Advanced Protein Crystallography Research Group, RIKEN Har
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NAKAYAMA Hiroshi
Biomolecular Characterization Team, Advanced Development and Supporting Center, RIKEN
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UEDA Shunsaku
United Graduate School of Agricultural Science, Tokyo University of Agriculture and Technology
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YOHDA Masafumi
Faculty of Technology, Tokyo University of Agriculture and Technology
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Kawano Yoshiaki
Protein Crystallography Methodology Team Advanced Protein Crystallography Research Group Riken Harim
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Takarada Hiromi
United Graduate School Of Agricultural Science Tokyo University Of Agriculture And Technology
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Ueda Shunsaku
United Graduate School Of Agricultural Science Tokyo University Of Agriculture And Technology
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Hashimoto Koichi
Department Of Biotechnology And Life Science Graduate School Of Engineering Tokyo University Of Agri
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Nakayama Hiroshi
Biomolecular Characterization Team Advanced Development And Supporting Center Riken
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Nakayama Hiroshi
Biomolecular Characterization Division Riken (the Institute Of Physical And Chemical Research)
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