PMID: 2115440Jul 5, 1990Paper

A 31P-nuclear-magnetic-resonance study of NADPH-cytochrome-P-450 reductase and of the Azotobacter flavodoxin/ferredoxin-NADP+ reductase complex

European Journal of Biochemistry
P J BonantsD E Edmondson

Abstract

31P-nuclear-magnetic-resonance spectroscopy has been employed to probe the structure of the detergent-solubilized form of liver microsomal NADPH--cytochrome-P-450 reductase. In addition to the resonances due to the FMN and FAD coenzymes, additional phosphorus resonances are observed and are assigned to the tightly bound adenosine 2'-phosphate (2'-AMP) and to phospholipids. The phospholipid content was found to vary with the preparation; however, the 2'-AMP resonance was observed in all preparations tested. In agreement with published results [Otvos et al. (1986) Biochemistry 25, 7220-7228] for the protease-solubilized enzyme, the addition of Mn(II) to the oxidized enzyme did not result in any observable line-broadening of the FMN and FAD phosphorus resonances. The phospholipid resonances, however, were extensively broadened and the line width of the phosphorus resonance assigned to the bound 2'-AMP was broadened by approximately 70 Hz. The data show that only the phosphorus moieties of the phospholipids and the 2'-AMP, but not the flavin coenzymes are exposed to the bulk solvent. Removal of the FMN moiety from the enzyme substantially alters the 31P-NMR spectrum as compared with the native enzyme. The 2'-AMP is removed from the...Continue Reading

References

Apr 21, 1975·Biochemical and Biophysical Research Communications·J D Dignam, H W Strobel
Aug 1, 1979·Proceedings of the National Academy of Sciences of the United States of America·D E Edmondson, T L James
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Citations

Jun 19, 2010·ACS Chemical Biology·Alexis S IvanovLarissa M Podust
Nov 19, 2003·European Journal of Biochemistry·Marco H HeftiWillem J H van Berkel
Apr 11, 1995·Proceedings of the National Academy of Sciences of the United States of America·S DjordjevicJ J Kim
Aug 5, 1997·Proceedings of the National Academy of Sciences of the United States of America·M WangJ J Kim
Dec 15, 1993·European Journal of Biochemistry·W G BalversI M Rietjens
Apr 23, 1996·Biochemistry·A WillieM S Jorns

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