小鼠神经酰胺葡萄糖基转移酶的分子克隆与表达

Molecular cloning and expression of mouse ceramide glucosyltransferase.

作者信息

Ichikawa S, Ozawa K, Hirabayashi Y

机构信息

Laboratory for Cellular Glycobiology, The Institute of Chemical and Physical Research (RIKEN), Saitama, Japan.

出版信息

Biochem Mol Biol Int. 1998 May;44(6):1193-202. doi: 10.1080/15216549800202282.

DOI:10.1080/15216549800202282

PMID:9623774

Abstract

Ceramide glucosyltransferase (EC 2.4.1.80) catalyzes the first glycosylation step of glycosphingolipid (GSL) synthesis, the transfer of glucose from UDP-Glucose to hydrophobic ceramide and generate glucosylceramide (GlcCer). We have cloned mouse ceramide glucosyltransferase cDNA from a brain cDNA library by PCR based homology cloning. The nucleotide sequence determination revealed that mouse ceramide glucosyltransferase cDNA encodes 394 amino acids with a calculated molecular mass of 45 kDa. The amino acid sequence of mouse ceramide glucosyltransferase showed 98% identity with the human sequence. Homology searches against currently available databases identified three homologous proteins in Caenorhabditis elegans and one homologous protein in Cyanobacteria. Highly conserved sequences of ceramide glucosyltransferases and the homologs among a wide variety of organisms suggest biological significance of the lipid glucosylation system.

摘要

神经酰胺葡萄糖基转移酶（EC 2.4.1.80）催化糖鞘脂（GSL）合成的第一步糖基化反应，即将葡萄糖从尿苷二磷酸葡萄糖转移至疏水的神经酰胺上，生成葡萄糖神经酰胺（GlcCer）。我们通过基于PCR的同源克隆技术，从小鼠脑cDNA文库中克隆出了小鼠神经酰胺葡萄糖基转移酶cDNA。核苷酸序列测定结果显示，小鼠神经酰胺葡萄糖基转移酶cDNA编码394个氨基酸，计算所得分子量为45 kDa。小鼠神经酰胺葡萄糖基转移酶的氨基酸序列与人类序列的一致性为98%。对现有数据库进行同源性搜索发现，秀丽隐杆线虫中有三种同源蛋白，蓝细菌中有一种同源蛋白。神经酰胺葡萄糖基转移酶及其在多种生物中的同源物的高度保守序列表明了脂质糖基化系统的生物学意义。