Title | Facile Formation of -thioGlcNAc Linkages to Thiol-Containing Sugars, Peptides, and Proteins using a Mutant GH20 Hexosaminidase |
Publication Type | Journal Article |
Year of Publication | 2019 |
Authors | Tegl, G, Hanson, J, Chen, H-M, Kwan, DH, Santana, AG, Withers, SG |
Journal | ANGEWANDTE CHEMIE-INTERNATIONAL EDITION |
Volume | 58 |
Pagination | 1632-1637 |
Date Published | FEB 4 |
ISSN | 1433-7851 |
Abstract | Thioglycosides are hydrolase-resistant mimics of O-linked glycosides that can serve as valuable probes for studying the role of glycosides in biological processes. The development of an efficient, enzyme-mediated synthesis of thioglycosides, including S-GlcNAcylated proteins, is reported, using a thioglycoligase derived from a GH20 hexosaminidase from Streptomyces plicatus in which the catalytic acid/base glutamate has been mutated to an alanine (SpHex E314A). This robust, easily-prepared, engineered enzyme uses GlcNAc and GalNAc donors and couples them to a remarkably diverse set of thiol acceptors. Thioglycoligation using 3-, 4-, and 6-thiosugar acceptors from a variety of sugar families produces S-linked disaccharides in nearly quantitative yields. The set of possible thiol acceptors also includes cysteine-containing peptides and proteins, rendering this mutant enzyme a promising catalyst for the production of thio analogues of biologically important GlcNAcylated peptides and proteins. |
DOI | 10.1002/anie.201809928 |
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