EXPRESSION OF CATALYTICALLY ACTIVE RADISH 3-HYDROXY-3-METHYLGLUTARYL COENZYME-A REDUCTASE IN ESCHERICHIA-COLI

被引:36
作者
FERRER, A
APARICIO, C
NOGUES, N
WETTSTEIN, A
BACH, TJ
BORONAT, A
机构
[1] UNIV BARCELONA, FAC FARM, UNITAT BIOQUIM, PZA PIUS XII SN, E-08028 BARCELONA, SPAIN
[2] UNIV KARLSRUHE, INST BOT 2, W-7500 KARLSRUHE, GERMANY
来源
FEBS LETTERS | 1990年 / 266卷 / 1-2期
关键词
Cloned enzyme expression; Escherichia coli; HMG-CoA reductase; Isoprenoid biosynthesis; Rhaphanus sativus;
D O I
10.1016/0014-5793(90)81508-L
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
Two fragments of a cDNA encoding radish 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR) were cloned into the vector pET-8c and expressed in Escherichia coli. The large fragment, encoding both the membrane and the cytosolic domains, was expressed at low level, essentially as an insoluble protein without enzymatic activity. In contrast, the fragment encoding only the cytosolic domain was expressed at a high level in a catalytically active form. The amount of soluble active enzyme in cell-free extracts of E. coli dramatically increased when the temperature during the induction was lowered from 37°C to 22°C. © 1990.
引用
收藏
页码:67 / 71
页数:5
相关论文
共 25 条
[1]   MEVINOLIN - A HIGHLY POTENT COMPETITIVE INHIBITOR OF HYDROXYMETHYLGLUTARYL-COENZYME-A REDUCTASE AND A CHOLESTEROL-LOWERING AGENT [J].
ALBERTS, AW ;
CHEN, J ;
KURON, G ;
HUNT, V ;
HUFF, J ;
HOFFMAN, C ;
ROTHROCK, J ;
LOPEZ, M ;
JOSHUA, H ;
HARRIS, E ;
PATCHETT, A ;
MONAGHAN, R ;
CURRIE, S ;
STAPLEY, E ;
ALBERSSCHONBERG, G ;
HENSENS, O ;
HIRSHFIELD, J ;
HOOGSTEEN, K ;
LIESCH, J ;
SPRINGER, J .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES, 1980, 77 (07) :3957-3961
[2]  
BACH TJ, 1987, PLANT PHYSIOL BIOCH, V25, P163
[3]   DETERGENT-SOLUBILIZATION, PURIFICATION, AND CHARACTERIZATION OF MEMBRANE-BOUND 3-HYDROXY-3-METHYLGLUTARYL-COENZYME-A REDUCTASE FROM RADISH SEEDLINGS [J].
BACH, TJ ;
ROGERS, DH ;
RUDNEY, H .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 1986, 154 (01) :103-111
[4]   INHIBITION BY MEVINOLIN OF PLANT-GROWTH, STEROL FORMATION AND PIGMENT ACCUMULATION [J].
BACH, TJ ;
LICHTENTHALER, HK .
PHYSIOLOGIA PLANTARUM, 1983, 59 (01) :50-60
[5]  
BACH TJ, 1983, Z NATURFORSCH C, V38, P212
[6]  
BACH TJ, 1987, ACS SYM SER, V325, P109
[7]   INHIBITORS OF STEROL BIOSYNTHESIS AND GROWTH IN PLANTS AND FUNGI [J].
BURDEN, RS ;
COOKE, DT ;
CARTER, GA .
PHYTOCHEMISTRY, 1989, 28 (07) :1791-1804
[8]  
BURNETTE WN, 1981, ANAL BIOCHEM, V112, P195, DOI 10.1016/0003-2697(81)90281-5
[9]   ISOLATION AND STRUCTURAL CHARACTERIZATION OF A CDNA-ENCODING ARABIDOPSIS-THALIANA 3-HYDROXY-3-METHYLGLUTARYL COENZYME-A REDUCTASE [J].
CAELLES, C ;
FERRER, A ;
BALCELLS, L ;
HEGARDT, FG ;
BORONAT, A .
PLANT MOLECULAR BIOLOGY, 1989, 13 (06) :627-638
[10]  
GRAY JC, 1987, ADV BOT RES, V14, P25