Molecular cloning of cDNA for cholesterol 7 alpha-hydroxylase from rat liver microsomes. Nucleotide sequence and expression.
A complete cDNA clone encoding cholesterol 7 alpha-hydroxylase was isolated from a rat liver cDNA library by the use of specific antibodies to the enzyme. The isolated cDNA clone was 3.6 kbp long and contained a 1509-bp open reading frame encoding 503 amino acid residues (Mr = 56,880). The identity of the cDNA was confirmed by expression of cholesterol 7 alpha-hydroxylase activity and the immunoreactive protein in COS cells transfected with pSVL expression vector carrying the cDNA insert. The primary structure of cholesterol 7 alpha-hydroxylase deduced from the nucleotide sequence of the cDNA indicated that the enzyme constitutes a novel P-450 family.
Noshiro
M
M
Department of Biochemistry, School of Dentistry, Hiroshima University, Japan.
Nishimoto
M
M
Morohashi
K
K
Okuda
K
K
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D002460:Cell Line; D002790:Cholesterol 7-alpha-Hydroxylase / Q000235:genetics*; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics*; D015870:Gene Expression; D015723:Gene Library; D008862:Microsomes, Liver / Q000201:enzymology*; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D051381:Rats; D015183:Restriction Mapping; D013250:Steroid Hydroxylases / Q000235:genetics*; D014162:Transfection
D004247:DNA; D013250:Steroid Hydroxylases; D002790:Cholesterol 7-alpha-Hydroxylase