title: Conformational States of Cytochrome P450 Oxidoreductase Evaluated by Förster Resonance Energy Transfer Using Ultrafast Transient Absorption Spectroscopy.
abstract: NADPH-cytochrome P450 oxidoreductase (CYPOR) was shown to undergo large conformational rearrangements in its functional cycle. Using a new Förster resonance energy transfer (FRET) approach based on femtosecond transient absorption spectroscopy (TA), we determined the donor-acceptor distance distribution in the reduced and oxidized states of CYPOR. The unmatched time resolution of TA allowed the quantitative assessment of the donor-acceptor FRET, indicating that CYPOR assumes a closed conformation in both reduced and oxidized states in the absence of the redox partner. The described ultrafast TA measurements of FRET with readily available red-infrared fluorescent labels open new opportunities for structural studies in chromophore-rich proteins and their complexes.
authors.affiliation: Biochemistry Department, Medical College of Wisconsin , Milwaukee, Wisconsin 53226, United States.|Chemistry Department, Marquette University , P.O. Box 1881, Milwaukee, Wisconsin 53201, United States.
authors.forename: Elizaveta A
authors.initials: EA
authors.lastname: Kovrigina
authors.affiliation: Chemistry Department, Marquette University , P.O. Box 1881, Milwaukee, Wisconsin 53201, United States.
authors.forename: Brian
authors.initials: B
authors.lastname: Pattengale
authors.affiliation: Biochemistry Department, Medical College of Wisconsin , Milwaukee, Wisconsin 53226, United States.
authors.forename: Chuanwu
authors.initials: C
authors.lastname: Xia
authors.affiliation: Chemistry Department, Marquette University , P.O. Box 1881, Milwaukee, Wisconsin 53201, United States.
authors.forename: Azamat R
authors.initials: AR
authors.lastname: Galiakhmetov
authors.affiliation: Chemistry Department, Marquette University , P.O. Box 1881, Milwaukee, Wisconsin 53201, United States.
authors.forename: Jier
authors.initials: J
authors.lastname: Huang
authors.affiliation: Biochemistry Department, Medical College of Wisconsin , Milwaukee, Wisconsin 53226, United States.
authors.forename: Jung-Ja P
authors.initials: JP
authors.lastname: Kim
authors.affiliation: Chemistry Department, Marquette University , P.O. Box 1881, Milwaukee, Wisconsin 53201, United States.
authors.forename: Evgenii L
authors.initials: EL
authors.lastname: Kovrigin
mesh_terms: D018360:Crystallography, X-Ray; D031541:Fluorescence Resonance Energy Transfer; D009251:NADPH-Ferrihemoprotein Reductase / Q000737:chemistry*; D011487:Protein Conformation
chemical_list: D009251:NADPH-Ferrihemoprotein Reductase