Hydroxylase activity of Met471Cys tyramine beta-monooxygenase

Journal of the American Chemical Society
Corinna R HessJ P Klinman

Abstract

A series of mutations was targeted at the methionine residue, Met471, coordinating the Cu(M) site of tyramine beta-monooxygenase (TbetaM). The methionine ligand at Cu(M) is believed to be key to dioxygen activation and the hydroxylation chemistry of the copper monooxygenases. The reactivity and copper binding properties of three TbetaM mutants, Met471Asp, Met471Cys, and Met471His, were examined. All three mutants show similar metal binding affinities to wild type TbetaM in the oxidized enzyme forms. EPR spectroscopy suggests that the Cu(II) coordination geometry is identical to that of the WT enzyme. However, substrate hydroxylation was observed for the reaction of tyramine solely with Met471Cys TbetaM. Met471Cys TbetaM provides the first example of an active mutant directed at the Cu(M) site of this class of hydroxylases. The reactivity and altered kinetics of the Met471Cys mutant further highlight the central role of the methionine residue in the enzyme mechanism. The sole ability of the cysteine residue to support activity among the series of alternate amino acids investigated is relevant to theoretical and biomimetic investigations of dioxygen activation at mononuclear copper centers.

References

Jan 1, 1978·Methods in Enzymology·C WalshR Spencer
Mar 1, 1980·The Biochemical Journal·S G Waley
Jul 25, 2000·Journal of Biological Inorganic Chemistry : JBIC : a Publication of the Society of Biological Inorganic Chemistry·N J BlackburnS Jaron
Sep 5, 2003·Chemistry & Biology·Niroshini M GilesClaus Jacob
Dec 14, 2005·Journal of Biological Inorganic Chemistry : JBIC : a Publication of the Society of Biological Inorganic Chemistry·Benjamin F GhermanChristopher J Cramer
Feb 1, 2006·Journal of Biological Inorganic Chemistry : JBIC : a Publication of the Society of Biological Inorganic Chemistry·William B Tolman
Sep 22, 2006·Antioxidants & Redox Signaling·Janet V Cross, Dennis J Templeton
Apr 27, 2007·Accounts of Chemical Research·Christopher J Cramer, William B Tolman

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Citations

Jun 15, 2010·Journal of Biological Inorganic Chemistry : JBIC : a Publication of the Society of Biological Inorganic Chemistry·Corinna R HessNinian J Blackburn
Sep 6, 2013·Inorganic Chemistry·Tetsuro TanoShinobu Itoh
Jan 20, 2017·Chemical Reviews·Courtney E ElwellWilliam B Tolman
Sep 3, 2009·Angewandte Chemie·Grégory ThiabaudOlivia Reinaud
Aug 9, 2020·Angewandte Chemie·Meredith A AllenAndré M Beauchemin
Mar 9, 2021·Journal of the American Chemical Society·Mayukh BhadraKenneth D Karlin
Apr 11, 2020·Inorganic Chemistry·Sethuraman MuthuramalingamRamasamy Mayilmurugan

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