Isolation of a cDNA for adrenodoxin reductase (ferredoxin-NADP+ reductase). Implications for mitochondrial cytochrome P-450 systems.
Using specific antibodies against adrenodoxin reductase (AR), we screened lambda gt11 cDNA expression libraries constructed from bovine adrenal cortex mRNA, and isolated several putative clones coding for this enzyme. Concurrently we determined the amino acid sequences of fragments from it. A deoxyinosine-containing oligonucleotide probe, generated for one of the sequences, reacted specifically with one of the cloned cDNAs of about 1600 base pairs. The codon sequence of this cDNA matched the peptide sequences, further confirming its identity as a copy of AR mRNA. RNA blot analysis indicates that in the adrenal cortex and corpus luteum there is only one major mRNA (approximately 2000 bp) for AR. The levels of this mRNA are at least 40-fold lower in the liver and kidney which are also known to contain in homologue of AR. As compared to adrenodoxin and cytochrome P-450scc mRNAs, AR mRNA levels in the adrenal cortex appear to be about 10-fold lower. Southern blot analysis of bovine and human genomic DNAs reveals that in both of these species there is only one gene for AR. These results indicate that only a single reductase serves the different mitochondrial P-450 systems in steroidogenic tissues.
Hanukoglu
I
I
Department of Biology, Technion-Israel Institute of Technology, Haifa, Israel.
Gutfinger
T
T
Haniu
M
M
Shively
J E
JE
D000302:Adrenal Cortex / Q000201:enzymology; D000325:Adrenodoxin / Q000235:genetics; D000818:Animals; D002417:Cattle; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics*; D004247:DNA / Q000302:isolation & purification*; D005287:Ferredoxin-NADP Reductase / Q000235:genetics*; D005786:Gene Expression Regulation; D006801:Humans; D007527:Isoenzymes / Q000235:genetics; D008928:Mitochondria / Q000201:enzymology*; D009247:NADH, NADPH Oxidoreductases / Q000235:genetics*; D009693:Nucleic Acid Hybridization
D007527:Isoenzymes; D000325:Adrenodoxin; D004247:DNA; D003577:Cytochrome P-450 Enzyme System; D005287:Ferredoxin-NADP Reductase; D009247:NADH, NADPH Oxidoreductases