D88A mutant of cytochrome P450nor provides kinetic evidence for direct complex formation with electron donor NADH

European Journal of Biochemistry
Mariko UmemuraH Shoun

Abstract

The haem-distal pocket of nitric oxide reductase cytochrome P450 contains many Arg and Lys residues that are clustered to form a putative access channel for NADH. Asp88 is the sole negatively charged amino acid in this positive charge cluster, and thus it would be interesting to know its functional role. Here we found the intriguing phenomenon that mutation at this site of P450nor (D88A or D88V) considerably decreased the overall nitric oxide reductase activity without blocking the reducing half reaction in which the ferric enzyme-NO complex is reduced with NADH to yield a specific intermediate (I). The results indicate that the catalytic turnover subsequent to the I formation was blocked by such mutation. This property of the mutants made it possible to perform kinetic analysis of the reduction step, which is impossible with the wild-type P450nor. These results are the first kinetic evidence for direct complex formation between P450nor and an electron donor (NADH or NADPH). The kinetic analysis also showed that the inhibition by chloride ions (Cl(-)) is competitive with respect to NAD(P)H, which highlights the importance of the binding site for Cl(-) (the anion hole) in the interaction with NAD(P)H. We also characterized anoth...Continue Reading

References

Aug 27, 1999·Archives of Biochemistry and Biophysics·D R Nelson
Dec 22, 1999·Biochemical and Biophysical Research Communications·T Omura
Nov 7, 1996·Chemical Reviews·Masanori SonoJohn H. Dawson
Mar 19, 2002·Journal of Inorganic Biochemistry·A DaiberV Ullrich
Jul 10, 2002·The Journal of Biological Chemistry·Li ZhangHirofumi Shoun

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Citations

Apr 30, 2011·Chemical Reviews·Daniela MontiSergio Riva
Mar 28, 2012·Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences·Hirofumi ShounTakayoshi Wakagi
Jun 1, 2004·Bioscience, Biotechnology, and Biochemistry·Fei SuHirofumi Shoun
Aug 18, 2004·Journal of Molecular Biology·Rieko OshimaHirofumi Shoun
Mar 18, 2006·Journal of Molecular Graphics & Modelling·Dóra K Menyhárd, György M Keseru
Jan 14, 2010·Journal of Biochemistry·Tsuneo Omura
Nov 19, 2011·Chemphyschem : a European Journal of Chemical Physics and Physical Chemistry·Christoph Riplinger, Frank Neese
Feb 29, 2020·Chemical Reviews·Christina FerousiKyle M Lancaster
Jun 9, 2005·Chemical Reviews·Ilia G DenisovIlme Schlichting
May 19, 2021·Proceedings of the National Academy of Sciences of the United States of America·Takashi NomuraYoshitsugu Shiro

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