Sterol 27-hydroxylase: expression in human arterial endothelium.

Human endothelium obtained from both the aorta and the pulmonary artery has been evaluated for the presence of the messenger RNA coding for the expression of sterol 27-hydroxylase. Unique oligomers were designed to detect the mRNA by reverse transcription followed by the polymerase chain reaction. The amplified product was sequenced and was found to be identical to the published sequence for nucleotides 491 to 802 of the human sterol 27-hydroxylase cDNA. Northern blot analysis confirmed the presence of 27-hydroxylase mRNA in pulmonary artery and aortic endothelium. As part of these studies, enzymatic activity was assayed in cultured arterial endothelium using cholesterol as a substrate and isotope ratio gas-liquid chromatography-mass spectrometry to identify the metabolites, 27-hydroxycholesterol and 3 beta-hydroxy-5-cholestenoic acid, in the medium. Localization of sterol 27-hydroxylase to vascular endothelium indicates intracellular production of the biologically active metabolite 27-hydroxycholesterol.

[1]  D. Stocco,et al.  Role of the steroidogenic acute regulatory protein (StAR) in steroidogenesis. , 1996, Biochemical pharmacology.

[2]  W. Miller,et al.  Structure of the human steroidogenic acute regulatory protein (StAR) gene: StAR stimulates mitochondrial cholesterol 27-hydroxylase activity. , 1995, Biochemistry.

[3]  M. Waterman A rising StAR: an essential role in cholesterol transport. , 1995, Science.

[4]  O. Andersson,et al.  Atherosclerosis and sterol 27-hydroxylase: evidence for a role of this enzyme in elimination of cholesterol from human macrophages. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[5]  A. Galloway,et al.  Sterol 27-hydroxylase: high levels of activity in vascular endothelium. , 1994, Journal of lipid research.

[6]  N. Javitt,et al.  Cholesterol and 27-hydroxycholesterol 7 alpha-hydroxylation: evidence for two different enzymes. , 1993, Journal of lipid research.

[7]  N. Javitt,et al.  Bile acid and sterol solubilization in 2-hydroxypropyl-beta-cyclodextrin. , 1992, Journal of lipid research.

[8]  M. Osame,et al.  Atherogenic risk factors in cerebrotendinous xanthomatosis. , 1991, Clinica chimica acta; international journal of clinical chemistry.

[9]  U. Francke,et al.  Mutations in the bile acid biosynthetic enzyme sterol 27-hydroxylase underlie cerebrotendinous xanthomatosis. , 1991, The Journal of biological chemistry.

[10]  D. Russell,et al.  Characterization of human sterol 27-hydroxylase. A mitochondrial cytochrome P-450 that catalyzes multiple oxidation reaction in bile acid biosynthesis. , 1991, The Journal of biological chemistry.

[11]  E. Myers,et al.  Basic local alignment search tool. , 1990, Journal of molecular biology.

[12]  M. V. Doyle,et al.  Quantitation of mRNA by the polymerase chain reaction. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[13]  S. Shefer,et al.  Long-term treatment of cerebrotendinous xanthomatosis with chenodeoxycholic acid. , 1984, The New England journal of medicine.

[14]  K. Okuda,et al.  Hepatic mitochondrial cytochrome P-450: isolation and functional characterization. , 1977, Proceedings of the National Academy of Sciences of the United States of America.

[15]  R. Scheuermann,et al.  Polymerase chain reaction-based mRNA quantification using an internal standard: analysis of oncogene expression. , 1993, Methods in enzymology.