EN-BLOC INCORPORATION OF COATOMER SUBUNITS DURING THE ASSEMBLY OF COP-COATED VESICLES

被引:129
作者
HARAKUGE, S
KUGE, O
ORCI, L
AMHERDT, M
RAVAZZOLA, M
WIELAND, FT
ROTHMAN, JE
机构
[1] MEM SLOAN KETTERING CANC CTR, PROGRAM CELLULAR BIOCHEM & BIOPHYS, NEW YORK, NY 10021 USA
[2] UNIV HEIDELBERG, INST BIOCHEM 1, W-6900 HEIDELBERG, GERMANY
[3] UNIV GENEVA, SCH MED, INST HISTOL & EMBRYOL, CH-1211 GENEVA 4, SWITZERLAND
关键词
D O I
10.1083/jcb.124.6.883
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
The cDNA encoding epsilon-COP, the 36-kD subunit of coatomer, was cloned from a bovine liver cDNA library and sequenced. Immunoblotting with an anti-epsilon-COP antibody showed that epsilon-COP exists in COP-coated vesicles as well as in the cytosolic coatomer. Using the cloned cDNA, recombinant His(6)-tagged epsilon-COP was overexpressed in cultured Chinese hamster ovary (CHO) cells, from which metabolically radiolabeled coatomer was purified by taking advantage of the His(6) tag. Radiolabeled coatomer was employed to establish that all the subunits of the coatomer enter coated vesicles as an intact unit.
引用
收藏
页码:883 / 892
页数:10
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