title: NADPH-cytochrome P450 oxidoreductase from the chicken (Gallus gallus): sequence characterization, functional expression and kinetic study.
abstract: Cytochrome P450 monooxygenases have been well known to be responsible for the synthesis of endogenous compounds and the metabolism of exogenous compounds in almost all living organisms, which require NADPH-cytochrome P450 oxidoreductase (POR) as an electron donor to function. In this study, a 2031 bp open reading frame of POR gene was cloned from 35-day-old Roman hen liver, encoding an enzyme of 676 amino acids. Sequence analysis showed that chicken POR shares high homology with other vertebrates PORs and possesses the conserved binding domains of FAD, FMN, and NADPH. The genomic sequences of POR genes from chicken and other four vertebrates have highly conserved exon/intron organization structure. By fusion with bacterial signal peptide, chicken POR gene was functionally expressed in E. coli membrane and showed activities in reduction of cytochrome c and oxidation of NADPH. The Km values for cytochrome c and NADPH were 21.9 ± 2.3 μM and 2.4 ± 0.3 μM respectively. A Ping-Pong mechanism was proposed for chicken POR.
authors: State Key Laboratory of Integrated Management of Pest Insects and Rodents, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China.
authors: Xiaojie
authors: X
authors: Zhou
authors: Mei
authors: M
authors: Li
authors: Chengfa
authors: C
authors: Sheng
authors: Xinghui
authors: X
authors: Qiu
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002645:Chickens / Q000235:genetics / Q000378:metabolism*; D005260:Female; D007700:Kinetics; D008969:Molecular Sequence Data; D009251:NADPH-Ferrihemoprotein Reductase / Q000235:genetics / Q000378:metabolism*; D020539:Sequence Analysis, Protein
chemical_list: D009251:NADPH-Ferrihemoprotein Reductase