Cloning, expression, and functional characterization of a Ca2+-dependent endoplasmic reticulum nucleoside diphosphatase

被引：48

作者：

Failer, BU ^{[1
]}

Braun, N ^{[1
]}

Zimmermann, H ^{[1
]}

机构：

[1] Goethe Univ Frankfurt, Biozentrum, D-60439 Frankfurt, Germany

来源：

JOURNAL OF BIOLOGICAL CHEMISTRY | 2002年 / 277卷 / 40期

关键词：

D O I：

10.1074/jbc.M201656200

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：

We have isolated and characterized the cDNA encoding a Ca2+-dependent nucleoside diphosphatase (EC 3.6.1.6) related to two secreted ATP- and ADP-hydrolyzing apyrases of the bloodsucking insects, Cimex lectularius and Phlebotomus papatasi. The rat brain-derived cDNA has an open reading frame of 1209 bp encoding a protein of 403 amino acids and a calculated molecular mass of 45.7 kDa. The mRNA was expressed in all tissues investigated, revealing two major transcripts with varying preponderance. The immunohistochemical analysis of the Myc-His-tagged enzyme expressed in Chinese hamster ovary cells revealed its association with the endoplasmic reticulum And also with pre-Golgi intermediates. Ca2+-dependent nucleoside diphosphatase is a membrane protein with its catalytic site facing the organelle lumen. It hydrolyzes nucleoside 5'-diphosphates in the order UDP >GDP = IDP > > >CDP but not ADP. Nucleoside 5'-triphosphates were hydrolyzed to a minor extent, and no hydrolysis of nucleoside 5'-monophosphates was observed. The enzyme was strongly activated by Ca2+, insensitive to Mg2+, and had a K-m for UDP of 216 muM. Ca2+-dependent nucleoside diphosphatase may support glycosylation reactions related to quality control in the endoplasmic reticulum.

引用

页码：36978 / 36986

页数：9

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