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A systematic analysis of acceptor specificity and reaction kinetics of five human α(23)sialyltransferases: Product inhibition studies illustrates reaction mechanism for ST3Gal-I

机译:对五种人类α(23)唾液酸转移酶的受体特异性和反应动力学的系统分析:产品抑制研究阐明了ST3Gal-1的反应机理

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摘要

Sialyltransferases (STs) catalyze the addition of sialic acids to the non-reducing ends of glycoproteins and glycolipids. In this work, we examined the acceptor specificity of five human α(2,3)sialyltransferases, namely ST3Gal-I, -II, -III, -IV and -VI. KM values for each of these enzymes is presented using radioactivity for acceptors containing Type-I (Galβ1,3GlcNAc), Type-II (Galβ1,4GlcNAc), Type-III (Galβ1,3GalNAc) and Core-2 (Galβ1,3(GlcNAcβ1,6)GalNAc) reactive groups. Several variants of acceptors inhibited ST3Gal activity emphasizing structural role of acceptor in enzyme-catalyzed reactions. In some cases, mass spectrometry was performed for structural verification. The results demonstrate human ST3Gal-I catalysis towards Type-III and Core-2 acceptors with KM = 5–50μM and high VMax values. The KM for ST3Gal-I and ST3Gal-II was 100 and 30-fold lower, respectively, for Type-III compared to Type-I acceptors. Variants of Type-I and Type-II structures characterized ST3Gal-III, -IV and -VI for their catalytic specificity. This manuscript also estimates KM for human ST3Gal-VI using Type-I and Type-II substrates. Together, these findings built a platform for designing inhibitors of STs having therapeutic potential.
机译:唾液酸转移酶(ST)催化向糖蛋白和糖脂的非还原末端添加唾液酸。在这项工作中,我们检查了五个人类α(2,3)唾液酸转移酶,即ST3Gal-1,-II,-III,-IV和-VI的受体特异性。使用包含I型(Galβ1、3GlcNAc),II型(Galβ1、4GlcNAc),III型(Galβ1、3GalNAc)和Core-2(Galβ1、3(GlcNAcβ1) ,6)GalNAc)反应性基团。受体的几种变体抑制ST3Gal活性,从而强调了受体在酶催化反应中的结构作用。在某些情况下,使用质谱进行结构验证。结果表明,人ST3Gal-I催化KM =5-50μM和高VMax值的III型和Core-2型受体。与I型受体相比,III型的ST3Gal-1和ST3Gal-II的KM分别低100倍和30倍。 I型和II型结构的变体表征了ST3Gal-III,-IV和-VI的催化特异性。该手稿还使用I型和II型底物估算了人类ST3Gal-VI的KM。这些发现共同建立了一个平台,用于设计具有治疗潜力的STs抑制剂。

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