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Title: Transferase Versus Hydrolase: The Role of Conformational Flexibility in Reaction Specificity

Journal Article · · Structure
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Active in the aqueous cellular environment where a massive excess of water is perpetually present, enzymes that catalyze the transfer of an electrophile to a non-water nucleophile (transferases) require specific strategies to inhibit mechanistically related hydrolysis reactions. To identify principles that confer transferase versus hydrolase reaction specificity, we exploited two enzymes that use highly similar catalytic apparatuses to catalyze the transglycosylation (a transferase reaction) or hydrolysis of α-1,3-glucan linkages in the cyclic tetrasaccharide cycloalternan (CA). In this work we show that substrate binding to non-catalytic domains and a conformationally stable active site promote CA transglycosylation, whereas a distinct pattern of active site conformational change is associated with CA hydrolysis. These findings defy the classic view of induced-fit conformational change and illustrate a mechanism by which a stable hydrophobic binding site can favor transferase activity and disfavor hydrolysis. Application of these principles could facilitate the rational reengineering of transferases with desired catalytic properties.

Research Organization:
Argonne National Laboratory (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
Sponsoring Organization:
Michigan Economic Development Corporation and the Michigan Technology Tri-Corridor; National Institute of Allergy and Infectious Diseases (NIAID); National Institutes of Health (NIH); US Department of Health and Human Services (HHS); USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22)
Grant/Contract Number:
AC02-06CH11357
OSTI ID:
1419804
Journal Information:
Structure, Journal Name: Structure Journal Issue: 2 Vol. 25; ISSN 0969-2126
Publisher:
ElsevierCopyright Statement
Country of Publication:
United Kingdom
Language:
English

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Related Subjects

59 BASIC BIOLOGICAL SCIENCES
carbohydrate
hydrolase
non-catalytic binding
protein crystallography
surface binding sites
transferase
transglycosidase
transglycosylase