MOLECULAR-CLONING OF GUINEA-PIG CYP1A1 - COMPLETE PRIMARY STRUCTURE AND FAST MOBILITY OF EXPRESSED PROTEIN ON ELECTROPHORESIS

被引:12
作者
OHGIYA, S
ISHIZAKI, K
SHINRIKI, N
机构
[1] Government Industrial Development Laboratory-Hokkaido, Agency of Industrial Science and Technology, Sapporo
关键词
CYTOCHROME-P-450; CDNA CLONING; PRIMARY STRUCTURE; EXPRESSION; (GUINEA PIG); (YEAST);
D O I
10.1016/0167-4781(93)90150-C
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
Guinea pig CYP1A1 cDNA was isolated from a liver cDNA library of guinea pig treated with 3-methylcholanthrene. The cDNA, named GPc1, was 2674 bp long and contained an entire coding region for 516 amino acids. The amino acid sequence of guinea pig CYP1A1 shared 74-78% identity with those of the other mammalian CYP1A1s. RNA blot and immunoblot analyses revealed that CYP1A1 was constitutively expressed and was induced by 3-methylcholanthrene in guinea pig liver. On sodium dodecyl sulfate-polyacrylamide gel electrophoresis, guinea pig CYP1A1 expressed in yeast had a significantly smaller apparent molecular mass than expressed mouse CYP1A1. An alignment of the amino acid sequences of mammalian CYP1A1s demonstrated that guinea pig CYP1A1 was several residues shorter than the counterparts in the N-terminal region. Thus, to clarify the contribution of the N-terminal sequence of guinea pig CYP1A1 to the fast mobility on the electrophoresis, mouse-guinea pig chimeric CYP1A1 was prepared through cDNA-directed expression in yeast. The chimeric CYP1A1 protein had an intermediate molecular mass between mouse and guinea pig CYP1A1s indicating that the anomalous mobility of guinea pig CYP1A1 is in part due to the shortened N-terminal amino acid sequence of the protein.
引用
收藏
页码:237 / 244
页数:8
相关论文
共 49 条
[1]   GENETIC-DIFFERENCES IN THE INDUCTION OF ARYL-HYDROCARBON HYDROXYLASE AND ITS COMPONENTS BY 3-METHYLCHOLANTHRENE IN LIVER AND LUNG MICROSOMES AMONG 4 STRAINS OF GUINEA-PIGS [J].
ABE, T ;
WATANABE, M .
BIOCHEMICAL PHARMACOLOGY, 1982, 31 (11) :2077-2082
[2]  
ABE T, 1983, MOL PHARMACOL, V23, P258
[4]  
CARDY RH, 1980, CANCER RES, V40, P1879
[5]   AMINO-TERMINAL SEQUENCE AND STRUCTURE OF MONOCLONAL-ANTIBODY IMMUNOPURIFIED CYTOCHROMES-P-450 [J].
CHENG, KC ;
PARK, SS ;
KRUTZSCH, HC ;
GRANTHAM, PH ;
GELBOIN, HV ;
FRIEDMAN, FK .
BIOCHEMISTRY, 1986, 25 (09) :2397-2402
[6]  
GONZALEZ FJ, 1989, PHARMACOL REV, V40, P243
[7]   STRUCTURAL CHARACTERISTICS OF CYTOCHROME-P-450 - POSSIBLE LOCATION OF THE HEME-BINDING CYSTEINE IN DETERMINED AMINO-ACID-SEQUENCES [J].
GOTOH, O ;
TAGASHIRA, Y ;
IIZUKA, T ;
FUJIIKURIYAMA, Y .
JOURNAL OF BIOCHEMISTRY, 1983, 93 (03) :807-817
[8]   ESTIMATION OF ISOZYMES OF MICROSOMAL CYTOCHROME-P-450 IN RATS, RABBITS, AND HUMANS USING IMMUNOCHEMICAL STAINING COUPLED WITH SODIUM DODECYL-SULFATE POLYACRYLAMIDE-GEL ELECTROPHORESIS [J].
GUENGERICH, FP ;
WANG, P ;
DAVIDSON, NK .
BIOCHEMISTRY, 1982, 21 (07) :1698-1706
[9]   OXIDATION OF TOXIC AND CARCINOGENIC CHEMICALS BY HUMAN CYTOCHROME-P-450 ENZYMES [J].
GUENGERICH, FP ;
SHIMADA, T .
CHEMICAL RESEARCH IN TOXICOLOGY, 1991, 4 (04) :391-407
[10]   PURIFICATION AND CHARACTERIZATION OF LIVER MICROSOMAL CYTOCHROMES-P-450 - ELECTROPHORETIC, SPECTRAL, CATALYTIC, AND IMMUNOCHEMICAL PROPERTIES AND INDUCIBILITY OF 8 ISOZYMES ISOLATED FROM RATS TREATED WITH PHENOBARBITAL OR BETA-NAPHTHOFLAVONE [J].
GUENGERICH, FP ;
DANNAN, GA ;
WRIGHT, ST ;
MARTIN, MV ;
KAMINSKY, LS .
BIOCHEMISTRY, 1982, 21 (23) :6019-6030