Peroxidase Active Site Activity Assay

Duggan, Kelsey C.; Musee, Joel; Marnett, Lawrence J.

doi:10.1007/978-1-59745-364-6_6

Cited by 5 publications

(6 citation statements)

References 11 publications

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“…Table 10 shows that the 1% rice hull extract also induced a decrease in the enzyme myeloperoxidase, a representative neutrophil biomarker, in the mouse ear tissues from 270.81 U to 95.32 U, or a 65.62% reduction. The extent of decrease in this inflammation-associated enzyme 38 is similar to the above-mentioned three biomarkers. It is, therefore, likely that the inhibitory effect against myeloperoxidase is mechanistically related to the corresponding effects on the biomarkers.…”

Section: ' Results and Discussionsupporting

confidence: 75%

Composition of Liquid Rice Hull Smoke and Anti-Inflammatory Effects in Mice

Kim¹,

Yang²,

Kang

et al. 2011

J. Agric. Food Chem.

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A new liquid rice hull smoke extract with a smoky aroma and sugar-like odor prepared by pyrolysis of rice hulls followed by liquefaction of the resulting smoke contained 161 compounds characterized by GC/MS. Antioxidative, antiallergic, and anti-inflammatory activities of the extract were assessed in vitro and in vivo. At pH 5, the extract inhibited 1-diphenyl-2-picrylhydrazyl (DPPH) free radicals and suppressed nitric oxide (NO) and β-hexosaminidase releases from lipopolysaccharide (LPS)-induced RAW264.7 mouse macrophage leukemia cells and ionophore A23187-stimulated RBL-2H3 rat basophilic cells without significant cytotoxicity. 12-O-Tetradecanolylphorbol-13-acetate (TPA) was applied to the ears of CD-1 mice to induce inflammation (edema), which was accompanied by increases in a series of biomarkers. Topical application of 1% of the extract as well as feeding mice a standard diet with 1% extract for two weeks significantly reduced the expression of biomarkers associated with the TPA-induced inflammation. These include tumor necrosis factor-α (TNF-α), IL-1β, interleukin-1β (IL-1β), interleukin-6 (IL-6), leukotriene B(4) (LTB(4)), prostaglandin E(2) (PGE(2)), myeloperoxidase (MPO). These in vitro and in vivo findings demonstrate the potential value of rice hull smoke extract derived from a major agricultural byproduct to serve as a new biomaterial for the improvement of food quality and safety and the environment.

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Section: ' Results and Discussionsupporting

confidence: 75%

Composition of Liquid Rice Hull Smoke and Anti-Inflammatory Effects in Mice

Kim¹,

Yang²,

Kang

et al. 2011

J. Agric. Food Chem.

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“…Values for the peroxidase activity of Fg-cat are directly comparable to the reported peroxidase activity of mammalian cyclooxygenase enzymes in oxidation of ABTS [29]: with COX-1 and COX-2 using 15 S -HPETE as the oxidizing co-substrate, the enzymes exhibit turnover numbers of 51 – 120 s −1 , a Km for 15-HPETE of 8.5 – 9.9 μM, and kcat/Km values of 6 – 12 μM −1 .s −1 [29]. In comparison, the kcat/Km for Fg-cat in oxidizing ABTS using 13-HPOTE as co-substrate gave the slightly higher value of 22 μM −1 .s −1 (Figure 8).…”

Section: Discussionsupporting

confidence: 70%

A fungal catalase reacts selectively with the 13S fatty acid hydroperoxide products of the adjacent lipoxygenase gene and exhibits 13S-hydroperoxide-dependent peroxidase activity

Teder

Boeglin

Schneider

et al. 2017

Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biolog

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The genome of the fungal plant pathogen Fusarium graminearum harbors six catalases, one of which has the sequence characteristics of a fatty acid peroxide-metabolizing catalase. We cloned and expressed this hemoprotein (designated as Fg-cat) along with its immediate neighbor, a 13S-lipoxygenase (cf. Brodhun et al, PloS One, e64919, 2013) that we considered might supply a fatty acid hydroperoxide substrate. Indeed, Fg-cat reacts abruptly with the 13S-hydroperoxide of linoleic acid (13S-HPODE) with an initial rate of 700–1300 s−1. By comparison there was no reaction with 9R- or 9S-HPODEs and extremely weak reaction with 13R-HPODE (~0.5% of the rate with 13S-HPODE). Although we considered Fg-cat as a candidate for the allene oxide synthase of the jasmonate pathway in fungi, the main product formed from 13S-HPODE was identified by UV, MS, and NMR as 9-oxo-10E-12,13-cis-epoxy-octadecenoic acid (with no traces of AOS activity). The corresponding analog is formed from the 13S-hydroperoxide of α-linolenic acid along with novel diepoxy-ketones and two C13 aldehyde derivatives, the reaction mechanisms of which are proposed. In a peroxidase assay monitoring the oxidation of ABTS, Fg-cat exhibited robust activity (kcat 550 s−1) using the 13S-hydroperoxy-C18 fatty acids as the oxidizing co-substrate. There was no detectable peroxidase activity using the corresponding 9S-hydroperoxides, nor with t-butyl hydroperoxide, and very weak activity with H2O2 or cumene hydroperoxide at micromolar concentrations of Fg-cat. Fg-cat and the associated lipoxygenase gene are present together in fungal genera Fusarium, Metarhizium and Fonsecaea and appear to constitute a partnership for oxidations in fungal metabolism or defense.

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“…This reduction occurs at the expense of oxidation of one of endogenous amino-acid residues (eg, tyrosine residues of cyt c) [13]. Compound II drives the oxygenase half-reaction of the cycle leading to a protein radical-driven hydrogen abstraction from a substrate, formation of a carbon-centered lipid radical and, upon addition of molecular oxygen, a peroxyl radical and an oxygenated product [14]. Our previous work indicates that tyrosyl radicals (Tyr•) are likely candidates for this role in cyt c/CL catalyzed peroxidation of polyunsaturated CLs [15] and abstraction of hydrogen from hydroxy-groups of phenolic compounds (eg, etoposide) [11].…”

Section: Introductionmentioning

confidence: 99%

Topography of tyrosine residues and their involvement in peroxidation of polyunsaturated cardiolipin in cytochrome c/cardiolipin peroxidase complexes

Kapralov

Yanamala

Tyurina

et al. 2011

Biochimica et Biophysica Acta (BBA) - Biomembranes

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Formation of cytochrome c (cyt c)/cardiolipin (CL) peroxidase complex selective towards peroxidation of polyunsaturated CLs is a pre-requisite for mitochondrial membrane permeabilization. Tyrosine residues – via the generation of tyrosyl radicals (Tyr•) - are likely reactive intermediates of the peroxidase cycle leading to CL peroxidation. We used mutants of horse heart cyt c in which each of the four Tyr residues was substituted for Phe and assessed their contribution to the peroxidase catalysis. Tyr67Phe mutation was associated with a partial loss of the oxygenase function of the cyt c/CL complex and the lowest concentration of H2O2-induced Tyr radicals in electron paramagnetic resonance (EPR) spectra. Our MS experiments directly demonstrated decreased production of CL-hydroperoxides (CL-OOH) by Tyr67Phe mutant. Similarly, oxidation of a phenolic substrate, Amplex Red, was affected to a greater extent in Tyr67Phe than in three other mutants. Tyr67Phe mutant exerted high resistance to H2O2-induced oligomerization. Measurements of Tyr fluorescence, hetero-nuclear magnetic resonance (NMR) and computer simulations position Tyr67 in close proximity to the porphyrin ring heme iron and one of the two axial heme-iron ligand residues, Met80. Thus, the highly conserved Tyr67 is a likely electron-donor (radical acceptor) in the oxygenase half-reaction of the cyt c/CL peroxidase complex.

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Peroxidase Active Site Activity Assay

Cited by 5 publications

References 11 publications

Composition of Liquid Rice Hull Smoke and Anti-Inflammatory Effects in Mice

Composition of Liquid Rice Hull Smoke and Anti-Inflammatory Effects in Mice

A fungal catalase reacts selectively with the 13S fatty acid hydroperoxide products of the adjacent lipoxygenase gene and exhibits 13S-hydroperoxide-dependent peroxidase activity

Topography of tyrosine residues and their involvement in peroxidation of polyunsaturated cardiolipin in cytochrome c/cardiolipin peroxidase complexes

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