Mutational Analysis of Amino Acid Residues Involved in Catalytic Activity of a Family 18 Chitinase from Tulip Bulbs(Biochemistry & Molecular Biology)
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概要
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We expressed chitinase-1 (TBC-1) from tulip bulbs (Tulipa bakeri) in E. coli cells and used site-directed mutagenesis to identify amino acid residues essential for catalytic activity. Mutations at Glu-125 and Trp-251 completely abolished enzyme activity, and activity decreased with mutations at Asp-123 and Trp-172 when glycolchitin was the substrate. Activity changed with the mutations of Trp-251 to one of several amino acids with side-chains of little hydrophobicity, suggesting that hydrophobic interaction of Trp-251 is important for the activity. Molecular dynamics (MD) simulation analysis with hevamine as the model compound showed that the distance between Asp-123 and Glu-125 was extended by mutation of Trp-251. Kinetic studies of Trp-251-mutated chitinases confirmed these various phenomena. The results suggested that Glu-125 and Trp-251 are essential for enzyme activity and that Trp-251 had a direct role in ligand binding.
- 社団法人日本農芸化学会の論文
- 2003-02-23
著者
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Yanagita T
Japan Institute Of Oil & Fats Other Foods Inspection Foundation
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ASO yoichi
Laboratory of Protein Chemistry & Engineering, Kyushu University
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ISHIGURO Masatsune
Laboratory of Protein Chemistry and Engineering, Kyushu University Graduate School of Bioresource an
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Yanagita Teruyoshi
Japan Institute Of Oil & Fats Other Foods Inspection Foundation
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Aso Y
Institute Of Genetic Resources Graduate School Of Bioresources And Bioenvironmental Science Kyushu U
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Ohnuma Takeshi
Laboratory Of Protein Chemistry And Engineering Graduate School Of Bioresource And Environmental Sci
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Kuhara Satoru
Genetic Regulation Graduate School Of Bioresource And Bioenvironmental Sciences Kyushu University
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SUZUKAWA Keisuke
Laboratories of Protein Chemistry and Engineering
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YAMAGAMI Takeshi
Laboratories of Protein Chemistry and Engineering
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OHNUMA Takayuki
Laboratories of Protein Chemistry and Engineering
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HIRAKAWA Hideki
Genetic Regulation, Graduate School of Bioresource and Bioenvironmental Sciences, Kyushu University
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Ishiguro M
Laboratory Of Insect Genetic Resources Faculty Of Agriculture Kyushu University
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Ishihara M
Laboratory Of Protein Chemistry And Engineering Graduate School Of Bioresource And Environmental Sci
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Ishiguro Masatsune
Laboratory Of Protein Chemistry & Engineering Department Of Genetic Resources Technology Facultr
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Yamagami T
Laboratory Of Protein Chemistry And Engineering Graduate School Of Bioresource And Environmental Sci
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Hirakawa H
Genetic Regulation Graduate School Of Bioresource And Bioenvironmental Sciences Kyushu University
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Ishiguro Masatsune
Laboratory of Insect Genetic Resources, Faculty of Agriculture, Kyushu University
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ASO Yoichi
Laboratory of Genetic and Protein Engineering, Faculty of Agriculture, Kyushu University
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