Purification and characterization of equine testicular cytochrome P-450 aromatase: comparison with the human enzyme

Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology
S MoslemiJ L Gaillard

Abstract

Cytochrome P-450 aromatase was purified by five chromatographic steps from adult stallion testis. It was first separated from NADPH-cytochrome P-450 reductase (reductase) on omega-aminohexyl-Sepharose 4B then purified to homogeneity on concanavalin A-Sepharose 4B, hydroxyapatite-Sepharose 4B, DEAE-Sepharose CL-6B and on a second hydroxyapatite-Sepharose 4B. On the other hand, purifications of the equine testicular and rat liver reductases, which allowed the reconstitution of aromatase activity in vitro, were achieved for each species in one chromatographic step on an adenosine 2',5'-diphosphate-agarose affinity column. Analysis on SDS/PAGE indicated single bands with apparent molecular masses of 53, 82, and 80 kDa for purified equine testicular cytochrome P-450 aromatase (eAROM), equine testicular reductase and rat liver reductase respectively. eAROM shows a time- and concentration-dependent activity that was stable for at least 2 months when stored at -78 degrees C. It is a highly hydrophobic protein composed from 505 residues and direct sequencing of its N-terminal part showed good homology when compared with human aromatase. When deglycosylated by N-glycosidase-F the apparent molecular mass of eAROM was decreased from 53 to ...Continue Reading

References

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Citations

May 9, 2007·Archives of Environmental Contamination and Toxicology·N BenachourG E Séralini
Nov 6, 2001·Journal of Biochemical and Biophysical Methods·G Houen
Feb 20, 2003·Biochimica Et Biophysica Acta·Gilles Eric SeraliniSafa Moslemi
Jun 19, 2001·Molecular and Cellular Endocrinology·G Séralini, S Moslemi
Oct 21, 1999·The Journal of Steroid Biochemistry and Molecular Biology·P AuvrayS Rault
Jun 3, 2005·Environmental Health Perspectives·Sophie RichardGilles-Eric Seralini
Jun 4, 2005·Drug Metabolism Reviews·Mike AguiarBernard F Gibbs
Sep 30, 2008·The Journal of Steroid Biochemistry and Molecular Biology·Todd W MillerCarlos L Arteaga
Dec 25, 2007·Virchows Archiv : an International Journal of Pathology·Luisa BarzonGiorgio Palù

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