caa a22 4500 477135749 CHVBK 20180405111717.0 cr unu---uuuuu 170330e19970701xx s 000 0 eng 10.1023/A:1018313830207 doi (NATIONALLICENCE)springer-10.1023/A:1018313830207 1H, 15N and 13C NMR resonance assignment, secondary structure and global fold of the FMN-binding domain of human cytochrome P450 [Elektronische Daten] [Igor Barsukov, Sandeep Modi, Lu-Yun Lian, Kong Sze, Mark Paine, C. Wolf, Gordon Roberts] The FMN-binding domain of human NADPH-cytochrome P450 reductase,corresponding to exons 3-;7, has been expressed at high level in anactive form and labelled with 13C and 15N. Mostof the backbone and aliphatic side-chain 1H, 15Nand 13C resonances have been assigned using heteronucleardouble- and triple-resonance methods, together with a semiautomaticassignment strategy. The secondary structure as estimated from the chemicalshift index and NOE connectivities consists of six α-helices and fiveβ-strands. The global fold was deduced from the long-range NOEsunambiguously assigned in a 4D 13C-resolved HMQC-NOESY-HMQCspectrum. The fold is of the alternating α/β type, with the fiveβ-strands arranged into a parallel β-sheet. The secondarystructure and global fold are very similar to those of the bacterialflavodoxins, but the FMN-binding domain has an extra short helix in place ofa loop, and an extra helix at the N-terminus (leading to the membrane anchordomain in the intact P450 reductase). The experimental constraints werecombined with homology modelling to obtain a structure of the FMN-bindingdomain satisfying the observed NOE constraints. Chemical shift comparisonsshowed that the effects of FMN binding and of FMN reduction are largelylocalised at the binding site. Kluwer Academic Publishers, 1997 Cytochrome P450 reductase nationallicence Flavin mononucleotide nationallicence Triple resonance nationallicence Resonance assignment nationallicence Semiautomatic nationallicence Barsukov Igor Department of Biochemistry and Biological NMR Centre, University of Leicester, LE1 9HN, Leicester, U.K aut Modi Sandeep Department of Biochemistry and Biological NMR Centre, University of Leicester, LE1 9HN, Leicester, U.K aut Lian Lu-Yun Department of Biochemistry and Biological NMR Centre, University of Leicester, LE1 9HN, Leicester, U.K aut Sze Kong Department of Biochemistry and Biological NMR Centre, University of Leicester, LE1 9HN, Leicester, U.K aut Paine Mark Biomedical Research Centre, Ninewells Hospital and Medical School, University of Dundee, DD1 9SY, Dundee, U.K aut Wolf C. Biomedical Research Centre, Ninewells Hospital and Medical School, University of Dundee, DD1 9SY, Dundee, U.K aut Roberts Gordon Department of Biochemistry and Biological NMR Centre, University of Leicester, LE1 9HN, Leicester, U.K aut Journal of Biomolecular NMR Kluwer Academic Publishers 10/1(1997-07-01), 63-75 0925-2738 10:1<63 1997 10 10858 https://doi.org/10.1023/A:1018313830207 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1023/A:1018313830207 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Barsukov Igor Department of Biochemistry and Biological NMR Centre, University of Leicester, LE1 9HN, Leicester, U.K aut NATIONALLICENCE 700 1- Modi Sandeep Department of Biochemistry and Biological NMR Centre, University of Leicester, LE1 9HN, Leicester, U.K aut NATIONALLICENCE 700 1- Lian Lu-Yun Department of Biochemistry and Biological NMR Centre, University of Leicester, LE1 9HN, Leicester, U.K aut NATIONALLICENCE 700 1- Sze Kong Department of Biochemistry and Biological NMR Centre, University of Leicester, LE1 9HN, Leicester, U.K aut NATIONALLICENCE 700 1- Paine Mark Biomedical Research Centre, Ninewells Hospital and Medical School, University of Dundee, DD1 9SY, Dundee, U.K aut NATIONALLICENCE 700 1- Wolf C. Biomedical Research Centre, Ninewells Hospital and Medical School, University of Dundee, DD1 9SY, Dundee, U.K aut NATIONALLICENCE 700 1- Roberts Gordon Department of Biochemistry and Biological NMR Centre, University of Leicester, LE1 9HN, Leicester, U.K aut NATIONALLICENCE 773 0- Journal of Biomolecular NMR Kluwer Academic Publishers 10/1(1997-07-01), 63-75 0925-2738 10:1<63 1997 10 10858 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer