Three-dimensional structure of the catalytic domain of chitinase Al from Bacillus circulars WL-12 at a very high resolution

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概要

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• The mature form of chitinase Al from Bacillus circulars WL-12 (one of the family 18 glycosyl hydrolases) comprises an N-terminal catalytic domain, two fibronectin type III domains and a C-terminal chitin-binding domain. In the efforts for crystallizing each domain, combined domains and the intact mature form, we have obtained crystals of the catalytic domain which diffract to a truly atomic resolution (1.13Å). The catalytic domain consists of an (α/β)8-TIM-barrel. Two small β-domains attached on top of the TIM-barrel provide a deep cleft for substrate binding. Crystals of an inactivated (through Glu204Gln mutation) catalytic domain complexed with a heptameric N-acetyl glucosamme (7NAG (NI, NII, NIII, NIV, NV, NVI, NVII-hepta-acetyl-chitoheptaose which is a β-1, 4-linked oligosaccharide of N-acetylglucosamine with a polymerization degree of seven.), which is a true substrate) diffracted up to 1.5Å resolution. The bound 7NAG substrate showed a marked kink between the sugar units at positions -1 and +1. The proximity of Glu204 to the scissile site strongly supports the hitherto conceived notion that Glu204 plays a role of catalytic acid in the so-called“substrate-assisted catalysis”mechanism which is distinct from the general acidbase mechanism long accepted for lysozyme and related glycosyl hydrolases.

• 日本学士院の論文

著者

• WATANABE Takeshi

  Faculty of Engineering, Kansai University

• Nonaka Takamasa

  Department Of Bioengineering Nagaoka University Of Technology

• MITSUI Yukio

  Department of BioEngineering Nagaoka University of Technology

• MATSUMOTO Takuo

  Department of BioEngineering, Nagaoka University of Technology

• HASHIMOTO Masayuki

  Faculty of Agriculture, Niigata University

• Watanabe Takeshi

  Faculty Of Education And Human Sciences Yamanashi University

• NONAKA Takamasa

  Department of BioEngineering, Nagaoka University of Technology

• WATANABE Takeshi

  Faculty of Agriculture, Niigata University

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