Generation of novel chimeric LacdiNAcS by gene fusion of α-lactalbumin and β1,4-galactosyltransferase 1

被引：0
作者：
Do, Su-Il [1 ,2 ]
机构：
[1] Ajou Univ, Dept Life Sci, Suwon 443749, South Korea
[2] Ajou Univ, Dept Mol Sci & Technol, Lab Funct Glyc, Suwon 443749, South Korea
来源：
GLYCOCONJUGATE JOURNAL | 2009年 / 26卷 / 05期
关键词：
beta 1,4-GalT1; alpha-LA; LacdiNAc; c-LacdiNAcS; Lec8 CHO cells; CHINESE-HAMSTER OVARY; LINKED OLIGOSACCHARIDES; N-ACETYLGALACTOSAMINYLTRANSFERASE; SCHISTOSOMA-MANSONI; MOLECULAR-CLONING; UDP-GALACTOSE; EXPRESSION; BETA; PROTEIN; BETA-1,4-N-ACETYLGALACTOSAMINYLTRANSFERASE;
D O I：
10.1007/s10719-008-9208-7
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：
Novel chimeric lacdiNAc (GalNAc(beta 1-4)GlcNAc) synthase (c-LacdiNAcS) was generated by gene fusion of alpha-lactalbumin (alpha-LA) and beta 1,4-galactosyltransferase 1 (beta 1,4-GalT1). c-LacdiNAcS was expressed in Lec8 Chinese hamster ovary (Lec8 CHO) cells and exhibited N-acetylgalactosaminyltransferase (GalNAcT) activity in the absence of exogenous alpha-LA as well as other glycosyltransferase activities including lactose synthase (LacS), and beta 1,4-GalT. These glycosyltransferase activities of c-LacdiNAcS were compared to those activities induced in LacS system under the co-presence of bovine beta 1,4-GalT1 and alpha-LA, indicating that each domain of alpha-LA and beta 1,4-GalT1 on c-LacdiNAcS is not only folding correctly, but also interacting together. Furthermore, c-LacdiNAcS was found to be auto-lacdiNAcylated and can synthesize lacdiNAc structures on cellular glycoproteins, demonstrating that GalNAcT activity of c-LacdiNAcS is functional in Lec8 CHO cells.
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页码：567 / 575
页数：9
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