Studying non-covalent enzyme carbohydrate interactions by STD NMR

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• 作者：Lothar Brecker ; Alex ; ra Schwarz ; Christiane Goedl ; Regina Kratzer ; Catrin E. Tyl ; Bernd Nidetzky
• 关键词：Binding-only conditions ; Co-substrate binding ; Glycostructures transforming enzyme ; Point mutated enzyme ; STD NMR
• 刊名：Carbohydrate Research
• 出版年：2008
• 出版时间：11 August 2008
• 年：2008
• 卷：343
• 期：12
• 页码：2153-2161
• 全文大小：506 K

文摘

Saturation transfer difference NMR spectroscopy is used to study non-covalent interactions between four different glycostructure transforming enzymes and selected substrates and products. Resulting binding patterns represent a molecular basis of specific binding between ligands and biocatalysts. Substrate and product binding to Aspergillus fumigatus glycosidase and to Candida tenuis xylose reductase are determined under binding-only conditions. Measurement of STD effects in substrates and products over the course of enzymatic conversion provides additional information about ligand binding during reaction. Influences of co-substrates and co-enzymes in substrate binding are determined for Schizophyllum commune trehalose phosphorylase and C. tenuis xylose reductase, respectively. Differences between ligand binding to wild type enzyme and a corresponding mutant enzyme are shown for Corynebacterium callunae starch phosphorylase and its His-334→Gly mutant. The resulting binding patterns are discussed with respect to the possibility that ligands do not only bind in the productive mode.