首页> 美国卫生研究院文献>Wiley-Blackwell Online Open >Facile Formation of β‐thioGlcNAc Linkages to Thiol‐Containing Sugars Peptides and Proteins using a Mutant GH20 Hexosaminidase
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Facile Formation of β‐thioGlcNAc Linkages to Thiol‐Containing Sugars Peptides and Proteins using a Mutant GH20 Hexosaminidase

机译:使用突变的GH20己糖胺酶容易地形成β-thioGlcNAc与硫醇的糖肽和蛋白质的连接

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摘要

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.
机译:硫代糖苷是O-连接糖苷的耐水解酶模拟物,可作为研究糖苷在生物过程中作用的有价值的探针。据报道,使用源自链霉菌(Streptomyces plicatus)的GH20己糖胺酶的硫糖寡糖酶,其中催化酸/谷氨酸催化物已突变为丙氨酸(SpHex E314A),开发了一种有效的酶介导的硫糖苷(包括S-GlcNA酰化蛋白)合成方法。 )。这种健壮,易于制备的工程化酶使用GlcNAc和GalNAc供体,并将它们与多种多样的硫醇受体结合。使用来自各种糖家族的3、4和6硫糖受体进行硫糖基化可以产生几乎定量的S连接二糖。这套可能的硫醇受体还包括含半胱氨酸的肽和蛋白质,使该突变酶成为生产具有生物学重要性的GlcNAcylated肽和蛋白质的硫代类似物的有希望的催化剂。

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