Novel features of acceptor recognition by β-(1→4)-galactosyltransferase

被引:28
作者
Kajihara, Y
Kodama, H
Endo, T
Hashimoto, H
机构
[1] Tokyo Inst Technol, Fac Biosci & Biotechnol, Dept Life Sci, Midori Ku, Yokohama, Kanagawa 227, Japan
[2] Japan Tobacco Inc, Life Sci Res Lab, Midori Ku, Yokohama, Kanagawa 227, Japan
关键词
beta-(1 -> 4)-galactosyltransferase; acceptor specificity; galactosyl transfer; N-acetylglucosamine;
D O I
10.1016/S0008-6215(97)10093-3
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
In order to understand how beta-(1-->4)-galactosyltransferase recognizes its glycosyl acceptor, substrate specificities were investigated using synthetic 2-acetamido-2-deoxy-D-glucopyranose (N-acetylglucosamine) derivatives in which the 1-, 2-, 3-, 4-, and 6-positions were systematically substituted. The hydroxyl groups at the 3-, 4-, and 6-positions were substituted by fluoride, thiol or hydrogen. For modification of the 2-position, the acetamido group was converted to ethylamino-, N-methylacetamido- and acetyloxy groups. For the anomeric position, several sugar residues were introduced as the aglycon of N-acetylglucosaminide. Galactose transfer assay using synthetic N-acetylglucosamine derivatives indicated that both the acetamido group and the 4-hydroxyl group were essential for binding of N-acetylglucosamine toward the beta-(1-->4)-galactosyltransferase. The assay also showed that the N-acetylglucosamine having a large substitution at the 6-position can be recognized as an acceptor. It is suggested that in this case the bulky substitutent is positioned away from the catalytic site or out of enzyme. Since the 2-acetamido and the 4-hydroxyl group are essential for recognition, the side composed of the 2, 3, and 4-positions may face the acceptor-binding site. (C) 1998 Elsevier Science Ltd. All rights reserved.
引用
收藏
页码:361 / 378
页数:18
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