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

共 28 条

[1]

AMBROOK J, 1989, MOL CLONING LABORATO

[2]

Ausubel F, 1988, CURRENT PROTOCOLS MO

[3] HUMAN CELL-LINES, DERIVED FROM AHH-1 TK+/- HUMAN LYMPHOBLASTS, GENETICALLY-ENGINEERED FOR EXPRESSION OF CYTOCHROMES-P450 [J].

CRESPI, CL ;

LANGENBACH, R ;

PENMAN, BW .

TOXICOLOGY, 1993, 82 (1-3) :89-104

[4] STABLE EXPRESSION OF HUMAN CYTOCHROME-P4502E1 IN HEPG2 CELLS - CHARACTERIZATION OF CATALYTIC ACTIVITIES AND PRODUCTION OF REACTIVE OXYGEN INTERMEDIATES [J].

DAI, Y ;

RASHBASTEP, J ;

CEDERBAUM, AI .

BIOCHEMISTRY, 1993, 32 (27) :6928-6937

[5]

DING XX, 1986, MOL PHARMACOL, V30, P370

[6]

ELIASSON E, 1992, J BIOL CHEM, V267, P15765

[7] ETHANOL-INDUCIBLE CYTOCHROME P4502E1 - GENETIC-POLYMORPHISM, REGULATION, AND POSSIBLE ROLE IN THE ETIOLOGY OF ALCOHOL-INDUCED LIVER-DISEASE [J].

INGELMANSUNDBERG, M ;

JOHANSSON, I ;

YIN, H ;

TERELIUS, Y ;

ELIASSON, E ;

CLOT, P ;

ALBANO, E .

ALCOHOL, 1993, 10 (06) :447-452

[8]

INOUE S, 1992, J BIOL CHEM, V267, P9080

[9] CATALYTIC EFFICIENCY OF EXPRESSED AROMATASE FOLLOWING SITE-DIRECTED MUTAGENESIS [J].

KADOHAMA, N ;

ZHOU, DJ ;

CHEN, S ;

OSAWA, Y .

BIOCHIMICA ET BIOPHYSICA ACTA, 1993, 1163 (02) :195-200

[10]

KAUFMAN RJ, 1990, METHOD ENZYMOL, V185, P537, DOI 10.1016/0076-6879(90)85044-O

← 1 2 3 →