A role of the heme-7-propionate side chain in cytochrome P450cam as a gate for regulating the access of water molecules to the substrate-binding site

Journal of the American Chemical Society
Takashi HayashiShun Hirota

Abstract

Cytochrome P450cam is a heme-containing enzyme which catalyzes hydroxylation of d-camphor. The heme is bound in the heme pocket via noncovalent interactions, where two heme-propionate side chains interact with Arg, His, and/or Asp residues. To understand the role of the heme-7-propionate side chain, we prepared reconstituted P450cam with an artificial one-legged heme which has a methyl group at the position of the 7-propionate. Removal of 7-propionate dramatically decreases the d-camphor affinity by 3 orders of magnitude relative to that of the wild-type enzyme, and spectroscopic data indicate that 74% of the ferric P450cam exhibits a low-spin state owing to water molecule occupancy in the substrate-binding site under the normal assay conditions. Thus, the monooxygenase activity of the reconstituted protein is remarkably low due to the decrease in the rate of the first electron transfer from reduced putidaredoxin, whereas 87% of oxidized NADH was utilized to produce 5-hydroxy-d-camphor without any significant uncoupling reactions. X-ray structural analysis of the reconstituted enzyme reveals a novel water array extending from the substrate-binding site to bulk solvent through the position occupied by 7-propionate. This water ar...Continue Reading

References

Jun 5, 1987·Journal of Molecular Biology·T L PoulosA J Howard
Mar 18, 1997·Proceedings of the National Academy of Sciences of the United States of America·T I OpreaA E Garcia
Mar 4, 2000·Science·I SchlichtingS G Sligar
Nov 7, 1996·Chemical Reviews·Masanori SonoJohn H. Dawson
May 29, 2003·Proceedings of the National Academy of Sciences of the United States of America·Victor GuallarRichard A Friesner
Jul 9, 2004·Journal of the American Chemical Society·Victor Guallar, Richard A Friesner
Jun 9, 2005·Chemical Reviews·Ilia G DenisovIlme Schlichting
Jun 9, 2005·Chemical Reviews·Sason ShaikWalter Thiel
Jun 24, 2006·BMC Bioinformatics·Martin PetrekJirí Damborský
May 31, 2007·Natural Product Reports·Sabine SchneiderMax Paoli
Dec 20, 2007·Journal of the American Chemical Society·Katsuyoshi HaradaTakashi Hayashi

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Citations

Jul 24, 2012·Applied Microbiology and Biotechnology·Stephen G BellLuet-Lok Wong
Jul 20, 2012·Biochemical and Biophysical Research Communications·Brenda J Ramos-Santana, Juan López-Garriga
Aug 28, 2013·Biochimica Et Biophysica Acta·Katrin Linke, Felix M Ho
Dec 15, 2010·Chembiochem : a European Journal of Chemical Biology·Ming MaZihe Rao
Dec 15, 2015·Beilstein Journal of Organic Chemistry·Paul P KellySabine L Flitsch
Oct 2, 2020·Database : the Journal of Biological Databases and Curation·Hiroko X KondoYu Takano
May 25, 2013·Chemistry, an Asian Journal·Ying-Wu LinJiangyun Wang
Oct 20, 2012·Chemical Communications : Chem Comm·Koji OohoraTakashi Hayashi
Nov 6, 2014·Acta Crystallographica. Section D, Biological Crystallography·Konrad HerzogAnett Schallmey
Apr 15, 2010·The Journal of Physical Chemistry. B·Dan FishelovitchRuth Nussinov
Mar 26, 2014·Chemical Reviews·Fangting YuVincent L Pecoraro
Jan 12, 2019·Accounts of Chemical Research·Kshatresh Dutta Dubey, Sason Shaik
Nov 28, 2018·Journal of the American Chemical Society·Navjeet Ahalawat, Jagannath Mondal
Jan 24, 2019·Journal of the American Chemical Society·Alec H FollmerThomas L Poulos

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