CLONING OF CYP11B1 AND CYP11B2 FROM NORMAL HUMAN ADRENAL AND THEIR FUNCTIONAL EXPRESSION IN COS-7 AND V79 CHINESE-HAMSTER CELLS

被引:57
作者
DENNER, K
DOEHMER, J
BERNHARDT, R
机构
[1] MAX DELBRUCK CTR MOLEC MED,D-13125 BERLIN,GERMANY
[2] FREE UNIV BERLIN,DEPT CHEM,BERLIN,GERMANY
[3] TECH UNIV MUNICH,INST TOXICOL,MUNICH,GERMANY
关键词
D O I
10.3109/07435809509030460
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
The cDNAs of human 11 beta hyxdroxylase (CYP11B1) and aldosterone synthase (CYP11B2) were cloned from surgically removed normal human adrenal using polymerase chain reaction. The cloned cDNAs were transfected into COS-7 cells and steroid hydroxylase activity of the two cytochromes P450 expressed was determined in order to elucidate their the functional characteristics. Stable expression of human CYP11B1 or CYP11B2 was performed in V79 hamster cells. and confirmed by Southern blotting, Northern blotting, and enzymatic activity. Interestingly, recombinant V79 cells were able to support CYP11B1 and CYP11B2 dependent steroid conversion without additional heterologous expression of the corresponding electron donor system.
引用
收藏
页码:443 / 448
页数:6
相关论文
共 13 条
[1]   EXPRESSION OF RAT-LIVER VITAMIN-D3 25-HYDROXYLASE CDNA IN SACCHAROMYCES-CEREVISIAE [J].
AKIYOSHISHIBATA, M ;
USUI, E ;
SAKAKI, T ;
YABUSAKI, Y ;
NOSHIRO, M ;
OKUDA, K ;
OHKAWA, H .
FEBS LETTERS, 1991, 280 (02) :367-370
[2]   V79 CHINESE-HAMSTER CELLS GENETICALLY-ENGINEERED FOR CYTOCHROME-P450 AND THEIR USE IN MUTAGENICITY AND METABOLISM STUDIES [J].
DOEHMER, J .
TOXICOLOGY, 1993, 82 (1-3) :105-118
[3]   STABLE EXPRESSION OF RAT CYTOCHROME P-450IIB1 CDNA IN CHINESE-HAMSTER CELLS (V79) AND METABOLIC-ACTIVATION OF AFLATOXIN-B1 [J].
DOEHMER, J ;
DOGRA, S ;
FRIEDBERG, T ;
MONIER, S ;
ADESNIK, M ;
GLATT, H ;
OESCH, F .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1988, 85 (16) :5769-5773
[4]   DIFFERENT ISOZYMES OF MOUSE 11-BETA-HYDROXYLASE PRODUCE MINERALOCORTICOIDS AND GLUCOCORTICOIDS [J].
DOMALIK, LJ ;
CHAPLIN, DD ;
KIRKMAN, MS ;
WU, RC ;
LIU, WW ;
HOWARD, TA ;
SELDIN, MF ;
PARKER, KL .
MOLECULAR ENDOCRINOLOGY, 1991, 5 (12) :1853-1861
[5]   NEW TECHNIQUE FOR ASSAY OF INFECTIVITY OF HUMAN ADENOVIRUS 5 DNA [J].
GRAHAM, FL ;
VANDEREB, AJ .
VIROLOGY, 1973, 52 (02) :456-467
[6]   CLONING AND EXPRESSION OF A CDNA FOR HUMAN CYTOCHROME-P-450ALDO AS RELATED TO PRIMARY ALDOSTERONISM [J].
KAWAMOTO, T ;
MITSUUCHI, Y ;
OHNISHI, T ;
ICHIKAWA, Y ;
YOKOYAMA, Y ;
SUMIMOTO, H ;
TODA, K ;
MIYAHARA, K ;
KURIBAYASHI, I ;
NAKAO, K ;
HOSODA, K ;
YAMAMOTO, Y ;
IMURA, H ;
SHIZUTA, Y .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1990, 173 (01) :309-316
[7]   ROLE OF STEROID 11-BETA-HYDROXYLASE AND STEROID 18-HYDROXYLASE IN THE BIOSYNTHESIS OF GLUCOCORTICOIDS AND MINERALOCORTICOIDS IN HUMANS [J].
KAWAMOTO, T ;
MITSUUCHI, Y ;
TODA, K ;
YOKOYAMA, Y ;
MIYAHARA, K ;
MIURA, S ;
OHNISHI, T ;
ICHIKAWA, Y ;
NAKAO, K ;
IMURA, H ;
ULICK, S ;
SHIZUTA, Y .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1992, 89 (04) :1458-1462
[8]  
MORNET E, 1989, J BIOL CHEM, V264, P20961
[9]   ENZYMATIC-ACTIVITIES OF P-450(11-BETA)S EXPRESSED BY 2 CDNAS IN COS-7 CELLS [J].
MOROHASHI, K ;
NONAKA, Y ;
KIRITA, S ;
HATANO, O ;
TAKAKUSU, A ;
OKAMOTO, M ;
OMURA, T .
JOURNAL OF BIOCHEMISTRY, 1990, 107 (04) :635-640
[10]  
OGISHIMA T, 1989, J BIOL CHEM, V264, P10935