Dolichylpyrophosphate oligosaccharides: Large-scale isolation and evaluation as oligosaccharyltransferase substrates

被引：22

作者：

Gibbs, BS ^{[1
]}

Coward, JK ^{[1
]}

机构：

[1] Univ Michigan, Coll Pharm, Dept Chem, Interdept Program Med Chem, Ann Arbor, MI 48109 USA

来源：

BIOORGANIC & MEDICINAL CHEMISTRY | 1999年 / 7卷 / 03期

关键词：

dolichylpyrophosphate; oligosaccharides; glycosylation; oligosaccharyltransferase;

D O I：

10.1016/S0968-0896(98)00268-5

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：

-Oligosaccharyltransferase (OST) catalyzes the transfer of a branched oligosaccharide from a dolichylpyrophosphate oligosaccharide (Dol-PP-OS) to the asparagine of a nascent polypeptide chain in vivo and peptide substrates in vitro. Here we report the isolation and purification of Dol-PP-Os from bovine pancreas and thyroid. Steady-state kinetic parameters comparing the two Dol-PP-OS to a shorter dolichylpyrophosphate disaccharide (DolPP-DS) previously synthesized in our laboratory are reported. These were determined for Dol-PP-OS, Dol-PP-DS, and the tripeptide Bz-Asn-Leu-Thr-NH2 with solubilized OST and, for the first time, saturation kinetics were observed for all substrates. The kinetic data provide a basis for analyzing quantitatively the individual contributions of oligosaccharide donor and peptide acceptor substrates to OST-catalyzed glycosylation. (C) 1999 Elsevier Science Ltd. All rights reserved.

引用

页码：441 / 447

页数：7

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