title: Expression of a recombinant derivative of human aromatase P450 in insect cells utilizing the baculovirus vector system.
abstract: Aromatase P450 (P450arom) is the enzyme responsible for estrogen biosynthesis. Studies of the relationship of the function of this enzyme to its structure have been hampered by lack of a suitable preparation. In the present report we describe the expression of a recombinant derivative of P450arom in insect cells by means of the baculovirus vector system. This protein, which lacks the first 41 amino acids from the N-terminus, and hence the membrane-spanning region, has spectral properties and activity similar to that of the wildtype protein. Moreover, the presence of a hexameric histidine tag at the C-terminus permits its facile purification by means of nickel-agarose affinity chromatography. This system permits the synthesis of quantities of a biologically active derivative of P450arom suitable for studies designed to explore the relationship of function to structure.
authors.affiliation: Cecil H and Ida Green Center for Reproductive Biology Sciences, University of Texas Southwestern Medical Center, Dallas, USA
authors.forename: B
authors.initials: B
authors.lastname: Amarneh
authors.forename: E R
authors.initials: ER
authors.lastname: Simpson
mesh_terms: D000735:Androstenedione / Q000378:metabolism; D000818:Animals; D001141:Aromatase / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D002460:Cell Line; D002846:Chromatography, Affinity; D004790:Enzyme Induction; D006801:Humans; D017924:Nucleopolyhedroviruses / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000302:isolation & purification; D018411:Spodoptera
chemical_list: D011993:Recombinant Fusion Proteins; D000735:Androstenedione; D001141:Aromatase