EXPRESSION OF AN ACTIVE GLYCOSYLATED HUMAN GAMMA-GLUTAMYL-TRANSPEPTIDASE MUTANT THAT LACKS A MEMBRANE ANCHOR DOMAIN

被引:49
作者
IKEDA, Y [1 ]
FUJII, J [1 ]
TANIGUCHI, N [1 ]
MEISTER, A [1 ]
机构
[1] OSAKA UNIV,SCH MED,DEPT BIOCHEM,SUITA,OSAKA 565,JAPAN
关键词
GAMMA-GLUTAMYL-TRANSFERASE; GLUTATHIONE; ENZYME PROCESSING; SIGNAL PEPTIDE;
D O I
10.1073/pnas.92.1.126
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
A mutant of human gamma-glutamyl transpeptidase (EC 2.3.2.2, a membrane-bound enzyme of importance in glutathione metabolism) that differs from the wild type by deletion of the putative signal peptide/anchor domain (amino acid residues 1-27) was expressed in insect cells using a baculovirus system. In contrast to the wild-type enzyme-which, as expected, was mainly cell-associated-the mutant enzyme was secreted into the medium. The mutant and wild-type enzymes were purified and found to exhibit virtually identical catalytic properties. The mutant enzyme was glycosylated and processed into two subunits, as found for the wild-type enzyme. Brefeldin A inhibited secretion of the mutant enzyme and led to its accumulation in cells. The findings indicate that gamma-glutamyl transpeptidase can be targeted to the endoplasmic reticulum in a manner that does not involve function of an amino-terminal ''signal/anchor'' domain and that this domain is involved primarily in a membrane anchoring function. Another region of the enzyme may function as a signal domain.
引用
收藏
页码:126 / 130
页数:5
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