title: [Overexpression, homology modeling and coenzyme docking studies of the cytochrome P450nor2 from Cylindrocarpon tonkinense].
abstract: Cytochrome P450nor catalyzes an unusual reaction that transfers electrons from NADP/NADPH to bound heme directly. To improve the expression level of P450nor2 from Cylindrocarpon tonkinense (C.P450nor2), Escherichia coli system was utilized to substitute the yeast system we constructed for expression of the P450nor2 gene, and the protein was purified in soluble form using Ni(+)-NTA affinity chromatography. In contrast to P450nor from Fusarium oxysporum (F.P450nor) and P450nor1 from Cylindrocarpon tonkinense (C.P450nor1), C.P450nor2 shows a dual specificity for using NADH or NADPH as electron donors. The present study developed a computational approach in order to illustrate the coenzyme specificity of C.P450nor2 for NADH and NADPH. This study involved homology modeling of C.P450nor2 and docking analyses of NADH and NADPH into the crystal structure of F.P450nor and the predictive model of C.P450nor2, respectively. The results suggested that C.P450nor2 and F.P450nor have different coenzyme specificity for NADH and NADPH; whilst the space around the B'-helix of the C.P450nor2, especially the Ser79 and Gly81, play a crucial role for the specificity of C.P450nor2. In the absence of the experimental structure of C.P450nor2, we hope that our model will be useful to provide rational explanation on coenzyme specificity of C.P450nor2.
authors.affiliation: Hubei Key Laboratory of Genetic Regulation and Integrative Biology, School of Life Science, Central China Normal University, Wuhan, 430079, China.|Key Laboratory of Crops with High Quality and Efficient Cultivation and Security Control, Yunnan Higher Education Institutions, College of Life Science and Technology, Honghe University, Mengzi, 661100, China.
authors.forename: N
authors.initials: N
authors.lastname: Li
authors.affiliation: Hubei Key Laboratory of Genetic Regulation and Integrative Biology, School of Life Science, Central China Normal University, Wuhan, 430079, China.
authors.forename: Y Z
authors.initials: YZ
authors.lastname: Zhang
authors.affiliation: Hubei Key Laboratory of Genetic Regulation and Integrative Biology, School of Life Science, Central China Normal University, Wuhan, 430079, China.
authors.forename: D D
authors.initials: DD
authors.lastname: Li
authors.affiliation: Hubei Key Laboratory of Genetic Regulation and Integrative Biology, School of Life Science, Central China Normal University, Wuhan, 430079, China.
authors.forename: Y H
authors.initials: YH
authors.lastname: Niu
authors.affiliation: Hubei Key Laboratory of Genetic Regulation and Integrative Biology, School of Life Science, Central China Normal University, Wuhan, 430079, China.
authors.forename: J
authors.initials: J
authors.lastname: Liu
authors.affiliation: Hubei Key Laboratory of Genetic Regulation and Integrative Biology, School of Life Science, Central China Normal University, Wuhan, 430079, China.
authors.forename: S X
authors.initials: SX
authors.lastname: Li
authors.affiliation: Hubei Key Laboratory of Genetic Regulation and Integrative Biology, School of Life Science, Central China Normal University, Wuhan, 430079, China.
authors.forename: Y Z
authors.initials: YZ
authors.lastname: Yuan
authors.affiliation: Hubei Key Laboratory of Genetic Regulation and Integrative Biology, School of Life Science, Central China Normal University, Wuhan, 430079, China.
authors.forename: S L
authors.initials: SL
authors.lastname: Chen
authors.affiliation: Hubei Key Laboratory of Genetic Regulation and Integrative Biology, School of Life Science, Central China Normal University, Wuhan, 430079, China.
authors.forename: H
authors.initials: H
authors.lastname: Geng
authors.affiliation: Hubei Key Laboratory of Genetic Regulation and Integrative Biology, School of Life Science, Central China Normal University, Wuhan, 430079, China.|ldl@mail.ccnu.edu.cn.
authors.forename: D L
authors.initials: DL
authors.lastname: Liu
mesh_terms: D002384:Catalysis; D003067:Coenzymes; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics; D005670:Fusarium / Q000201:enzymology*; D007700:Kinetics; D062105:Molecular Docking Simulation; D009249:NADP / Q000737:chemistry
chemical_list: D003067:Coenzymes; D009249:NADP; D003577:Cytochrome P-450 Enzyme System; C080610:nitric-oxide reductase (P450)
keywords: C.P450nor2; coenzyme specificity; homology modeling; molecular docking; overexpression