Expanding the thioglycoligase strategy to the synthesis of alpha-linked thioglycosides allows structural investigation of the parent enzyme/substrate complex.
For the first time, the thioglycoligase strategy has been successfully applied to alpha-glycosidases. The alpha-thioglycoligases derived from the family 31 glycosidases, alpha-xylosidase from E. coli (YicI) and alpha-glucosidase from Sulfolobus solfataricus, catalyze thioglycoligase reactions using alpha-glycosyl fluorides and deoxythioglycosides as donors and acceptors, respectively, in yields up to 86%. In addition, we describe the Michaelis complex of YicI using one of the thioglycosides as a nonhydrolyzable substrate analogue and discuss the structural insights this yields into the specificity and mechanism of family 31 alpha-glycosidases and the molecular basis of an associated genetic disease.
Kim
Young-Wan
YW
Department of Chemistry, University of British Columbia, 2036 Main Mall, Vancouver, British Columbia V6T 1Z1, Canada.
Lovering
Andrew L
AL
Chen
Hongming
H
Kantner
Terrence
T
McIntosh
Lawrence P
LP
Strynadka
Natalie C J
NC
Withers
Stephen G
SG
D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000201:enzymology; D029968:Escherichia coli Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry; D006026:Glycoside Hydrolases / Q000037:antagonists & inhibitors / Q000737:chemistry*; D008958:Models, Molecular; D048229:Sulfolobus solfataricus / Q000201:enzymology; D013865:Thioglycosides / Q000138:chemical synthesis* / Q000737:chemistry
D029968:Escherichia coli Proteins; D013865:Thioglycosides; C503200:YicI protein, E coli; D006026:Glycoside Hydrolases