RABBIT P450 2E1-EXPRESSED IN CHO-K1 CELLS HAS A SHORT HALF-LIFE

被引：25

作者：

BARMADA, S ^{[1
]}

KIENLE, E ^{[1
]}

KOOP, DR ^{[1
]}

机构：

[1] OREGON HLTH SCI UNIV, DEPT PHARMACOL, PORTLAND, OR 97206 USA

来源：

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS | 1995年 / 206卷 / 02期

关键词：

D O I：

10.1006/bbrc.1995.1085

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：

Rabbit P450 2E1 was stably expressed in Chinese hamster ovary cells after cotransfection with pRC/CMV-2E1 and pFR400 which expresses murine dihydrofolate reductase with a single arginine to leucine substitution at position 22. This mutation permits amplification of expression with increasing methotrexate concentrations in CHO-K1 cells that are not dihydrofolate reductase deficient. After amplification with 1 mu M methotrexate, a representative clone expressed about 15 pmol of P450 2E1/mg microsomal protein. Cells from a single 35-mm plate catalyzed the formation of 1.02 nmol 6-hydroxychlorzoxazone/10(6) cells/h or about 127 pmol/mg total cell protein/min. The enzyme was rapidly labeled when pulsed with [S-35]-methionine. Initial pulse-chase experiments indicate that the expressed protein has a half-life of 4.8 h. (C) 1995 Academic Press, Inc.

引用

页码：601 / 607

页数：7

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