Cloning, structure, and expression of the mitochondrial cytochrome P-450 sterol 26-hydroxylase, a bile acid biosynthetic enzyme

S. Anderson, D. L. Davis, H. Dahlback, H. Jornvall, D. W. Russell

Research output: Contribution to journalArticle

956 Citations (Scopus)

Abstract

The conversion of cholesterol into bile acids in the liver represents the major catabolic pathway for the removal of cholesterol from the body. In this complex biosynthetic pathway, at least 10 enzymes modify both the ring structure and side chain of cholesterol, resulting in the formation of the primary bile acids, cholic acid, and chenodeoxycholic acid. To gain insight into the details and regulation of this pathway, we have used protein sequencing and molecular cloning techniques to isolate and characterize a cDNA encoding the rabbit mitochondrial sterol 26-hydroxylase. This enzyme catalyzes the first step in the oxidation of the side chain of sterol intermediates in the biosynthesis of bile acids. The structure of the sterol 26-hydroxylase, as deduced by both DNA sequence analysis of the cDNA and protein sequence analysis, reveals it to be a mitochondrial cytochrome P-450. A signal sequence of 36 residues precedes a coding region of 499 amino acids, predicting a molecular weight of 56,657 for the mature protein. The identity of the 26-hydroxylase cDNA was further confirmed by expression in monkey COS cells employing a versatile eukaryotic expression vector. Blotting experiments revealed that the mRNA for this enzyme is expressed in many tissues and that it is encoded by a low copy number gene in the rabbit genome.

Original languageEnglish (US)
Pages (from-to)8222-8229
Number of pages8
JournalJournal of Biological Chemistry
Volume264
Issue number14
StatePublished - 1989

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Cholestanetriol 26-Monooxygenase
Cloning
Bile Acids and Salts
Organism Cloning
Complementary DNA
Cholesterol
Protein Sequence Analysis
Enzymes
Genes
Rabbits
Chenodeoxycholic Acid
Cholic Acid
Proteins
Gene Dosage
COS Cells
DNA sequences
Biosynthesis
Biosynthetic Pathways
Molecular Cloning
Sterols

ASJC Scopus subject areas

  • Biochemistry

Cite this

Cloning, structure, and expression of the mitochondrial cytochrome P-450 sterol 26-hydroxylase, a bile acid biosynthetic enzyme. / Anderson, S.; Davis, D. L.; Dahlback, H.; Jornvall, H.; Russell, D. W.

In: Journal of Biological Chemistry, Vol. 264, No. 14, 1989, p. 8222-8229.

Research output: Contribution to journalArticle

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AU - Jornvall, H.

AU - Russell, D. W.

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AB - The conversion of cholesterol into bile acids in the liver represents the major catabolic pathway for the removal of cholesterol from the body. In this complex biosynthetic pathway, at least 10 enzymes modify both the ring structure and side chain of cholesterol, resulting in the formation of the primary bile acids, cholic acid, and chenodeoxycholic acid. To gain insight into the details and regulation of this pathway, we have used protein sequencing and molecular cloning techniques to isolate and characterize a cDNA encoding the rabbit mitochondrial sterol 26-hydroxylase. This enzyme catalyzes the first step in the oxidation of the side chain of sterol intermediates in the biosynthesis of bile acids. The structure of the sterol 26-hydroxylase, as deduced by both DNA sequence analysis of the cDNA and protein sequence analysis, reveals it to be a mitochondrial cytochrome P-450. A signal sequence of 36 residues precedes a coding region of 499 amino acids, predicting a molecular weight of 56,657 for the mature protein. The identity of the 26-hydroxylase cDNA was further confirmed by expression in monkey COS cells employing a versatile eukaryotic expression vector. Blotting experiments revealed that the mRNA for this enzyme is expressed in many tissues and that it is encoded by a low copy number gene in the rabbit genome.

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