Effects of noncovalent and covalent FAD binding on the redox and catalytic properties of p-cresol methylhydroxylase

Biochemistry
Igor EfimovW McIntire

Abstract

Each flavoprotein subunit (alpha or PchF) of the alpha(2)beta(2) flavocytochrome p-cresol methylhydroxylase (PCMH) from Pseudomonas putida contains FAD covalently attached to Tyr384. PCMH oxidizes p-cresol to 4-hydroxybenzyl alcohol, which is oxidized subsequently by PCMH to 4-hydroxybenzaldehyde. The Y384F mutant form of PchF (apo-PchF[Y384F]) displayed stoichiometric noncovalent FAD binding. PchF[Y384F]FAD associated with the cytochrome subunit (beta or PchC) (producing PCMH[Y384F]), although not as avidly as with wild-type PchF containing covalently bound FAD (PchF(C)). Dramatic increases in the two-electron E(m,7) (NHE) values for FAD were observed when it bound noncovalently to either apo-PchF or apo-PchF[Y384F], and the two-electron E(m,7) value for FAD was increased further by about 75 mV upon covalent binding to PchF, i.e., PchF(C). The E(m,7) values increased by approximately 20 and 45 mV, respectively, when PchF(C) and PchF[Y384F]FAD associated with PchC. The two-electron E(m,7) for covalently bound FAD in PCMH is 84 mV, the highest measured for a flavoprotein. The values for the one-electron redox potentials (E(m,7), NHE) for FAD were measured also for various forms of PchF. Under anaerobiosis, the reduction of PchF[...Continue Reading

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Citations

Nov 19, 2003·European Journal of Biochemistry·Marco H HeftiWillem J H van Berkel
Sep 5, 2008·The Journal of Biological Chemistry·Chun-Hsiang HuangShwu-Huey Liaw
Apr 24, 2007·Journal of Bacteriology·Franziska PetersMatthias Boll
Oct 7, 2003·Annual Review of Biochemistry·Gary Cecchini
Nov 1, 2015·The FEBS Journal·David KopečnýSolange Moréra
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May 15, 2009·The FEBS Journal·Dominic P H M HeutsMarco W Fraaije
Sep 17, 2008·The FEBS Journal·Jianfeng JinMarco W Fraaije
Jan 24, 2009·Angewandte Chemie·Ljiljana FrukChristof M Niemeyer
Sep 21, 2006·The Journal of Biological Chemistry·Magdalena KujawaChristina Divne
Apr 1, 2007·EcoSal Plus·Thomas M TomasiakTina M Iverson

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