TY - JOUR
T1 - Saturation-transfer-difference NMR to characterize substrate binding recognition and catalysis of two broadly specific glycoside hydrolases
AU - Brecker, Lothar
AU - Straganz, Grit D.
AU - Tyl, Catrin E.
AU - Steiner, Walter
AU - Nidetzky, Bernd
N1 - Copyright:
Copyright 2008 Elsevier B.V., All rights reserved.
PY - 2006/11/2
Y1 - 2006/11/2
N2 - Saturation-transfer-difference NMR spectroscopy (STD NMR) is used to delineate noncovalent enzyme-substrate interactions of β-glycosidases from Pyrococcus furiosus and Aspergillus fumigatus under binding-only conditions at low temperatures, and during catalysis. Glucopyranosyl and galactopyranosyl moieties display a distinct pattern of multiple contacts with each active site, revealing enzyme-specific elements of recognition and portraying the global binding effect caused by single-site modification of the substrate, at carbon 4. The glucopyranose leaving group of cellobiose or lactose shows small relative STD effects except for the anomeric carbon, particularly in the α-form. Its replacement in β-glucosides by an alcohol leaving group strongly affects sugar binding in the proximal enzyme subsite. A combination of STD effects of substrate and product, produced by the catalytic event or added exogenously, characterizes subsite binding during cellobiose hydrolysis.
AB - Saturation-transfer-difference NMR spectroscopy (STD NMR) is used to delineate noncovalent enzyme-substrate interactions of β-glycosidases from Pyrococcus furiosus and Aspergillus fumigatus under binding-only conditions at low temperatures, and during catalysis. Glucopyranosyl and galactopyranosyl moieties display a distinct pattern of multiple contacts with each active site, revealing enzyme-specific elements of recognition and portraying the global binding effect caused by single-site modification of the substrate, at carbon 4. The glucopyranose leaving group of cellobiose or lactose shows small relative STD effects except for the anomeric carbon, particularly in the α-form. Its replacement in β-glucosides by an alcohol leaving group strongly affects sugar binding in the proximal enzyme subsite. A combination of STD effects of substrate and product, produced by the catalytic event or added exogenously, characterizes subsite binding during cellobiose hydrolysis.
KW - Enzyme-substrate interaction
KW - Glycoside hydrolysis
KW - Noncovalent binding recognition
KW - STD NMR
KW - β-Glycosidase
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U2 - 10.1016/j.molcatb.2006.07.004
DO - 10.1016/j.molcatb.2006.07.004
M3 - Article
AN - SCOPUS:33749237092
SN - 1381-1177
VL - 42
SP - 85
EP - 89
JO - Journal of Molecular Catalysis B: Enzymatic
JF - Journal of Molecular Catalysis B: Enzymatic
IS - 3-4
ER -