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Insights into the mechanism of Drosophila melanogaster Golgi alpha-mannosidase II through the structural analysis of covalent reaction intermediates

TitleInsights into the mechanism of Drosophila melanogaster Golgi alpha-mannosidase II through the structural analysis of covalent reaction intermediates
Publication TypeJournal Article
Year of Publication2003
AuthorsNumao, S, Kuntz, DA, Withers, SG, ROSE, DR
JournalJOURNAL OF BIOLOGICAL CHEMISTRY
Volume278
Pagination48074-48083
Date PublishedNOV 28
ISSN0021-9258
Abstract

The family 38 golgi alpha-mannosidase II, thought to cleave mannosidic bonds through a double displacement mechanism involving a reaction intermediate, is a clinically important enzyme involved in glycoprotein processing. The structure of three different covalent glycosyl-enzyme intermediates have been determined to 1.2-Angstrom resolution for the Golgi alpha-mannosidase II from Drosophila melanogaster by use of fluorinated sugar analogues, both with the wild-type enzyme and a mutant enzyme in which the acid/base catalyst has been removed. All these structures reveal sugar intermediates bound in a distorted S-1(5) skew boat conformation. The similarity of this conformation with that of the substrate in the recently determined structure of the Michaelis complex of a beta-mannanase (Ducros, V. M. A., Zechel, D. L., Murshudov, G. N., Gilbert, H. J., Szabo, L., Stoll, D., Withers, S. G., and Davies, G. J. (2002) Angew. Chem. Int. Ed. Engl. 41, 2824 - 2827) suggests that these disparate enzymes have recruited common stereoelectronic features in evolving their catalytic mechanisms.

DOI10.1074/jbc.M309249200