Insights into the catalytic mechanism of Grimontia hollisae collagenase through structural and mutational analyses.

Autor: Ueshima S; Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Japan., Yasumoto M; Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Japan., Kitagawa Y; Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Japan., Akazawa K; Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Japan., Takita T; Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Japan., Tanaka K; Nippi Research Institute of Biomatrix, Toride, Japan., Hattori S; Nippi Research Institute of Biomatrix, Toride, Japan., Mizutani K; Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Uji, Japan., Mikami B; Research Institute for Sustainable Humanosphere, Kyoto University, Uji, Japan.; Institute of Advanced Energy, Kyoto University, Uji, Japan., Yasukawa K; Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Japan.
Jazyk: angličtina
Zdroj: FEBS letters [FEBS Lett] 2023 Oct; Vol. 597 (19), pp. 2473-2483. Date of Electronic Publication: 2023 Sep 25.
DOI: 10.1002/1873-3468.14732
Abstrakt: Grimontia hollisae collagenase (Ghcol) exhibits high collagen-degrading activity. To explore its catalytic mechanism, its substrate (Gly-Pro-Hyp-Gly-Pro-Hyp, GPOGPO)-complexed crystal structure was determined at 2.0 Å resolution. A water molecule was observed near the active-site zinc ion. Since this water was not observed in the product (GPO)-complexed Ghcol, it was hypothesized that the GPOGPO-complexed Ghcol structure reflects a Michaelis complex, providing a structural basis for understanding the catalytic mechanism. Analyses of the active-site geometry and site-directed mutagenesis of the active-site tyrosine residues revealed that Glu493 and Tyr564 were essential for catalysis, suggesting that Glu493 functions as an acid and base catalyst while Tyr564 stabilizes the tetrahedral complex in the transition state. These results shed light on the catalytic mechanism of bacterial collagenase.
(© 2023 Federation of European Biochemical Societies.)
Databáze: MEDLINE
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