A Stable Bacterial Peroxidase with Novel Halogenating Activity and an Autocatalytically Linked Heme Prosthetic Group

Auer, Markus; Gruber, Clemens; Bellei, Marzia; Pirker, Katharina F.; Zámocký, Marcel; Kroiss, Daniela; Teufer, Stefan A.; Hofbauer, Stefan; Soudi, Monika; Battistuzzi, Gianantonio; Furtmüller, Paul G.; Obinger, Christian

doi:10.1074/jbc.m113.477067

Cited by 19 publications

(44 citation statements)

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“…Human peroxidases, like lactoperoxidase, eosinophil peroxidase and thyroid peroxidase autocatalytically form covalent haeme to protein ester linkages [50], whereas in myeloperoxidase an additional sulfonium linkage between a haeme vinyl group and a methionine is found [51]. Recently, it was shown that also some bacterial peroxidases establish covalent haeme to protein ester bonds [52,53]. However, in most haeme proteins the Fe-proximal His represents the only covalent bond between the haeme chromophore and the protein, and stable incorporation of the prosthetic group occurs via several non-covalent interactions.…”

Section: Discussionmentioning

confidence: 99%

From chlorite dismutase towards HemQ–the role of the proximal H-bonding network in haeme binding

et al. 2016

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SynopsisChlorite dismutase (Cld) and HemQ are structurally and phylogenetically closely related haeme enzymes differing fundamentally in their enzymatic properties. Clds are able to convert chlorite into chloride and dioxygen, whereas HemQ is proposed to be involved in the haeme b synthesis of Gram-positive bacteria. A striking difference between these protein families concerns the proximal haeme cavity architecture. The pronounced H-bonding network in Cld, which includes the proximal ligand histidine and fully conserved glutamate and lysine residues, is missing in HemQ. In order to understand the functional consequences of this clearly evident difference, specific hydrogen bonds in Cld from 'Candidatus Nitrospira defluvii' (NdCld) were disrupted by mutagenesis. The resulting variants (E210A and K141E) were analysed by a broad set of spectroscopic (UV-vis, EPR and resonance Raman), calorimetric and kinetic methods. It is demonstrated that the haeme cavity architecture in these protein families is very susceptible to modification at the proximal site. The observed consequences of such structural variations include a significant decrease in thermal stability and also affinity between haeme b and the protein, a partial collapse of the distal cavity accompanied by an increased percentage of low-spin state for the E210A variant, lowered enzymatic activity concomitant with higher susceptibility to self-inactivation. The high-spin (HS) ligand fluoride is shown to exhibit a stabilizing effect and partially restore wild-type Cld structure and function. The data are discussed with respect to known structure-function relationships of Clds and the proposed function of HemQ as a coprohaeme decarboxylase in the last step of haeme biosynthesis in Firmicutes and Actinobacteria.

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Section: Discussionmentioning

confidence: 99%

From chlorite dismutase towards HemQ–the role of the proximal H-bonding network in haeme binding

et al. 2016

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“…For ECD, Chirascan from Applied Photophysics (Leatherhead, UK) was used, and changes of the ellipticity at 208 nm as well as at 412 nm in 20 mM phosphate buffer, pH 7.4, with increasing temperature were recorded and evaluated (29). The instrument was flushed with nitrogen at a flow rate of 5 liters min Ϫ1 and was equipped with a Peltier element for temperature control.…”

Section: Mass Spectrometric Analysis Of Cysteines and Cystines-mentioning

confidence: 99%

Multidomain Human Peroxidasin 1 Is a Highly Glycosylated and Stable Homotrimeric High Spin Ferric Peroxidase

Soudi

Paumann-Page

Delporte

et al. 2015

Journal of Biological Chemistry

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Background: Human peroxidasin 1 (hsPxd01) mediates the formation of sulfilimine cross-links within the collagen IV scaffold of basement membranes. Results: Overexpressed hsPxd01 contains covalently linked heme catalytically active for production of hypobromous acid. Conclusion: hsPxd01 has peroxidase-like active site structure but restricted substrate accessibility. Significance: Architecture of hsPxd01 facilitates product release and its interactions with the physiological substrate collagen IV.

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“…These proteins have sequence similarity to animal heme peroxidases of the peroxidase-cyclooxygenase superfamily of peroxidases (18). This class of enzymes includes myeloperoxidases (MPO) and lactoperoxidases (LPO), which use hydrogen peroxide as the oxidant and, as their names suggest, contain a heme cofactor.…”

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“…This class of enzymes includes myeloperoxidases (MPO) and lactoperoxidases (LPO), which use hydrogen peroxide as the oxidant and, as their names suggest, contain a heme cofactor. These animal heme peroxidases generally consist of the peroxidase domain and another nonenzymatic domain (18,19). The Mn-oxidizing proteins (MopAs) of this type have been identified in Erythrobacter sp.…”

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confidence: 99%

“…strain SD21 and Aurantimonas manganoxydans strain SI85-9A1 by protein sequencing of Mn-oxidizing SDS-PAGE bands (5). These Mn-oxidizing proteins are in the peroxicin subfamily of the peroxidasecyclooxygenase superfamily (18,20). This subfamily has one or more animal heme peroxidase domains, with an additional domain containing multiple RTX-type calcium binding motifs (Fig.…”

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Heterologous Expression and Characterization of the Manganese-Oxidizing Protein from Erythrobacter sp. Strain SD21

Nakama

Medina

Lien

et al. 2014

Appl Environ Microbiol

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The manganese (Mn)-oxidizing protein (MopA) from Erythrobacter sp. strain SD21 is part of a unique enzymatic family that is capable of oxidizing soluble Mn(II). This enzyme contains two domains, an animal heme peroxidase domain, which contains the catalytic site, followed by a C-terminal calcium binding domain. Different from the bacterial Mn-oxidizing multicopper oxidase enzymes, little is known about MopA. To gain a better understanding of MopA and its role in Mn(II) oxidation, the 238-kDa fulllength protein and a 105-kDa truncated protein containing only the animal heme peroxidase domain were cloned and heterologously expressed in Escherichia coli. Despite having sequence similarity to a peroxidase, hydrogen peroxide did not stimulate activity, nor was activity significantly decreased in the presence of catalase. Both pyrroloquinoline quinone (PQQ) and hemin increased Mn-oxidizing activity, and calcium was required. The K m for Mn(II) of the full-length protein in cell extract was similar to that of the natively expressed protein, but the K m value for the truncated protein in cell extract was approximately 6-fold higher than that of the full-length protein, suggesting that the calcium binding domain may aid in binding Mn(II). Characterization of the heterologously expressed MopA has provided additional insight into the mechanism of bacterial Mn(II) oxidation, which will aid in understanding the role of MopA and Mn oxidation in bioremediation and biogeochemical cycling.

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A Stable Bacterial Peroxidase with Novel Halogenating Activity and an Autocatalytically Linked Heme Prosthetic Group

Cited by 19 publications

References 39 publications

From chlorite dismutase towards HemQ–the role of the proximal H-bonding network in haeme binding

From chlorite dismutase towards HemQ–the role of the proximal H-bonding network in haeme binding

Multidomain Human Peroxidasin 1 Is a Highly Glycosylated and Stable Homotrimeric High Spin Ferric Peroxidase

Heterologous Expression and Characterization of the Manganese-Oxidizing Protein from Erythrobacter sp. Strain SD21

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