BACULOVIRUS EXPRESSION OF BOVINE CYTOCHROME P450C17 IN SF9 CELLS AND COMPARISON WITH EXPRESSION IN YEAST, MAMMALIAN-CELLS, AND ESCHERICHIA-COLI

被引:28
作者
BARNES, HJ
JENKINS, CM
WATERMAN, MR
机构
[1] VANDERBILT UNIV,SCH MED,DEPT BIOCHEM,NASHVILLE,TN 37232
[2] UNIV TEXAS,SW MED CTR,DEPT BIOCHEM,DALLAS,TX 75235
关键词
D O I
10.1006/abbi.1994.1530
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
Expression of bovine 17 alpha-hydroxylase cytochrome P450 (P450c17) in insect Sf9 cells using baculovirus is the fourth heterologous expression system developed for studying this enzyme. The enzyme in Sf9 cell membranes has all the expected activities and closely resembles that from the other systems: COS1 cells, yeast, and Escherichia, coli. One curious feature of baculovirus expression of this hemoprotein is that the immunodetectable P450c17 is present in an insoluble pellet when produced in the absence of added hemin and in a 10,000g supernatant when expressed in the presence of added hemin. When comparing the level of expression of bovine P450c17 in these different expression systems, E. coli produces the largest quantity per liter culture, while baculovirus produces the most molecules of P450c17 per cell. (C) 1994 Academic Press, Inc.
引用
收藏
页码:489 / 494
页数:6
相关论文
共 23 条
[1]   NOVEL EXOGENOUS HEME-DEPENDENT EXPRESSION OF MAMMALIAN CYTOCHROME-P450 USING BACULOVIRUS [J].
ASSEFFA, A ;
SMITH, SJ ;
NAGATA, K ;
GILLETTE, J ;
GELBOIN, HV ;
GONZALEZ, FJ .
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1989, 274 (02) :481-490
[2]   EXPRESSION AND ENZYMATIC-ACTIVITY OF RECOMBINANT CYTOCHROME-P450 17-ALPHA-HYDROXYLASE IN ESCHERICHIA-COLI [J].
BARNES, HJ ;
ARLOTTO, MP ;
WATERMAN, MR .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1991, 88 (13) :5597-5601
[3]   CHARACTERIZATION OF COMPLEMENTARY DEOXYRIBONUCLEIC-ACID FOR HUMAN ADRENOCORTICAL 17-ALPHA-HYDROXYLASE - A PROBE FOR ANALYSIS OF 17-ALPHA-HYDROXYLASE DEFICIENCY [J].
BRADSHAW, KD ;
WATERMAN, MR ;
COUCH, RT ;
SIMPSON, ER ;
ZUBER, MX .
MOLECULAR ENDOCRINOLOGY, 1987, 1 (05) :348-354
[4]  
CLARK BJ, 1991, J BIOL CHEM, V266, P5898
[5]  
CLARK BJ, 1992, J BIOL CHEM, V267, P24568
[6]   RAT P45017-ALPHA FROM TESTIS - CHARACTERIZATION OF A FULL-LENGTH CDNA-ENCODING A UNIQUE STEROID HYDROXYLASE CAPABLE OF CATALYZING BOTH DELTA-4- AND DELTA-5-STEROID-17,20-LYASE REACTIONS [J].
FEVOLD, HR ;
LORENCE, MC ;
MCCARTHY, JL ;
TRANT, JM ;
KAGIMOTO, M ;
WATERMAN, MR ;
MASON, JI .
MOLECULAR ENDOCRINOLOGY, 1989, 3 (06) :968-975
[7]  
IMAI T, 1993, J BIOL CHEM, V268, P17317
[8]   DISSOCIATION OF HYDROXYLASE AND LYASE ACTIVITIES BY SITE-DIRECTED MUTAGENESIS OF THE RAT P45017-ALPHA [J].
KITAMURA, M ;
BUCZKO, E ;
DUFAU, ML .
MOLECULAR ENDOCRINOLOGY, 1991, 5 (10) :1373-1380
[9]   ALTERATION OF MOUSE CYTOCHROME-P450COH SUBSTRATE-SPECIFICITY BY MUTATION OF A SINGLE AMINO-ACID RESIDUE [J].
LINDBERG, RLP ;
NEGISHI, M .
NATURE, 1989, 339 (6226) :632-634
[10]   INCORPORATION OF STEROIDOGENIC PATHWAYS WHICH PRODUCE CORTISOL AND ALDOSTERONE FROM CHOLESTEROL INTO NONSTEROIDOGENIC CELLS [J].
MATHEW, PA ;
MASON, JI ;
TRANT, JM ;
WATERMAN, MR .
MOLECULAR AND CELLULAR ENDOCRINOLOGY, 1990, 73 (01) :73-80