Differences in the spectral interactions between NADPH-cytochrome P-450 reductase and a series of cytochrome P-450 enzymes

Biochem Biophys Res Commun. 1986 May 29;137(1):437-42. doi: 10.1016/0006-291x(86)91229-5.


The interaction between NADPH-cytochrome P-450 reductase and a series of cytochrome P-450 isozymes was investigated using UV-visible spectrophotometry. In the absence of substrate the interactions between the reductase and RLM3, RLM5, and RLM5a were tight, exhibiting sub-micromolar dissociation constants and resulted in type I spectra of varying magnitude from which the following increases in the proportion of high spin hemoprotein were calculated; RLM3 (7%), RLM5 (36%), RLM5a (6%), LM2 (29%), RLM2 (0%). Preincubation of LM2 with its type I substrate benzphetamine increased the affinity of the cytochrome for the reductase. Using initial estimates of the P-450 spin states in the absence of reductase in conjunction with the spectral binding data and equations relating these parameters to the microequilibria for the association of reductase with high or low spin P-450, RLM3, RLM5, RLM5a and LM2 were shown to bind significantly more tightly to high spin P-450. The relevance of this data to the understanding of spin state influence on P-450 reduction is discussed.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Cytochrome P-450 Enzyme System* / metabolism
  • Kinetics
  • Microsomes, Liver / enzymology
  • NADPH-Ferrihemoprotein Reductase* / metabolism
  • Protein Binding
  • Protein Conformation
  • Rabbits
  • Rats
  • Spectrum Analysis
  • Substrate Specificity


  • Cytochrome P-450 Enzyme System
  • NADPH-Ferrihemoprotein Reductase