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• W2998565526 abstract "A defining characteristic of bacterial cytochromes (cyt’s) in the P460 family is an unusual cross-link connecting the heme porphyrin to the side chain of a lysyl residue in the protein backbone. Here, via proteomics of the periplasmic fraction of the ammonia-oxidizing bacterium (AOB) Nitrosomonas europaea, we report the identification of a variant member of the P460 family that contains a methionyl residue in place of the cross-linking lysine. We formally designate this protein cytochrome “c′β-Met” to distinguish it from other members bearing different residues at this position (e.g., cyt c′β-Phe from the methane-oxidizing Methylococcus capsulatus Bath). As isolated, the monoheme cyt c′β-Met is high-spin (S = 5/2). Optical spectroscopy suggests that a cross-link is absent. Hydroxylamine, the substrate for the cross-linked cyt P460 from N. europaea, did not appreciably alter the optical spectrum of cyt c′β with up to 1000-fold excess at pH 7.5. Cyt c′β-Met did however bind 1 equiv of H2O2, and with a slight excess, Mössbauer spectroscopy indicated the formation of a semistable ferryl (FeIV═O) Compound II-like species. The corresponding electron paramagnetic resonance showed a very low intensity signal indicative of a radical at g = 2.0. Furthermore, cyt c′β-Met exhibited guaiacol-dependent peroxidase activity (kcat = 20.0 ± 1.2 s–1; KM = 2.6 ± 0.4 mM). Unlike cyt c′β-Met, cyt P460 showed evidence of heme inactivation in the presence of 2 equiv of H2O2 with no appreciable guaiacol-dependent peroxidase activity. Mutagenesis of the cross-linking lysyl residue to an alanine in cyt P460, however, reversed this lack of activity." @default.
• W2998565526 created "2020-01-10" @default.
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• W2998565526 date "2019-12-30" @default.
• W2998565526 modified "2023-09-23" @default.
• W2998565526 title "Cytochrome <i>c</i>′_β-Met Is a Variant in the P460 Superfamily Lacking the Heme–Lysyl Cross-Link: A Peroxidase Mimic Generating a Ferryl Intermediate" @default.
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• W2998565526 doi "https://doi.org/10.1021/acs.biochem.9b00810" @default.
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• W2998565526 hasPublicationYear "2019" @default.
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