Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:1.11.1.7 (peroxidase)
 65,474 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

In vivo most extracellular iron is bound to transferrin or lactoferrin in such a way as to be unable to catalyze the formation of hydroxyl radical from superoxide (.O2-) and hydrogen peroxide (H2O2). At sites of Pseudomonas aeruginosa infection bacterial and neutrophil products could possibly modify transferrin and/or lactoferrin forming catalytic iron complexes. To examine this possibility, diferrictransferrin and diferriclactoferrin which had been incubated with pseudomonas elastase, pseudomonas alkaline protease, human neutrophil elastase, trypsin, or the myeloperoxidase product HOCl were added to a hypoxanthine/xanthine oxidase .O2-/H2O2 generating system. Hydroxyl radical formation was only detected with pseudomonas elastase treated diferrictransferrin and, to a much lesser extent, diferriclactoferrin. This effect was enhanced by the combination of pseudomonas elastase with other proteases, most prominently neutrophil elastase. Addition of pseudomonas elastase-treated diferrictransferrin to stimulated neutrophils also resulted in hydroxyl radical generation. Incubation of pseudomonas elastase with transferrin which had been selectively iron loaded at either the NH2- or COOH-terminal binding site yielded iron chelates with similar efficacy for hydroxyl radical catalysis. Pseudomonas elastase and HOCl treatment also decreased the ability of apotransferrin to inhibit hydroxyl radical formation by a Fe-NTA supplemented hypoxanthine/xanthine oxidase system. However, apotransferrin could be protected from the effects of HOCl if bicarbonate anion was present during the incubation. Apolactoferrin inhibition of hydroxyl radical generation was unaffected by any of the four proteases or HOCl. Alteration of transferrin by enzymes and oxidants present at sites of pseudomonas and other bacterial infections may increase the potential for local hydroxyl radical generation thereby contributing to tissue injury.
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PMID:Pseudomonas and neutrophil products modify transferrin and lactoferrin to create conditions that favor hydroxyl radical formation. 165 25

The secondary structure of Pseudomonas cytochrome c peroxidase (ferrocytochrome c: hydrogen-peroxide oxidoreductase, EC 1.11.1.5) has been predicted from the established amino acid sequence of the enzyme using a Chou-Fasman-type algorithm. The amount of alpha-helicity thus obtained is in agreement with previously obtained results based on circular dichroic measurements at far UV. The two heme c moieties of the enzyme have earlier been shown to have widely different characteristics, e.g., the redox potentials of the hemes differ with about 600 mV, and carry out different functions in the enzyme molecule. The structural comparisons made in this study enlighten the observed functional differences. The first heme in the polypeptide chain, heme 1, has in its environment a folding pattern generally encountered in cytochromes. In the region of the sixth ligand, however, profound differences are noted. The cytochromal methionine has been replaced by a lysine with a concomitant lowering of redox-potential thus making peroxidatic activity possible. Around heme 2, extra amino acid residues have been added to the peroxidase as compared with Rhodospirillum molischianum cytochrome c2 core structure in the 20's loop. After completion of the cytochromal fold around heme 2 an additional tail consisting of 25 residues is linked. This tail shows no stabilizing elements of secondary structure, but contains a strongly hydrophobic segment which suggests a possible membrane contact site of this extrinsic membrane protein. Heme 2 is concluded to have a cytochromal function in the molecule. To further elucidate the functional properties of the enzyme, a noncovalent two-fragment complex was produced by specific cleavage of the peroxidase by Pseudomonas elastase. The complex was studied with respect to its properties to the native enzyme. The two-fragment complex of Pseudomonas peroxidase retains the overall conformation of the native enzyme showing, however, no heme-heme interaction. Thus, a comparison of the properties of the native enzyme with those of the two-fragment complex permitted some conclusions to be drawn on the structure of the enzyme as well as the mechanism of heme-heme interaction. From the present results we conclude that the two distal heme surfaces in the peroxidase are oriented toward each other. This structural arrangement allows an inter-heme communication in the enzyme molecule and it also forms the structural basis for the enzyme mechanism. The structural comparisons also give insight into the evolution of an ancestral cytochrome c into an efficient peroxidase that has a versatile control mechanism in heme-heme interaction.
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PMID:Structural and functional features of Pseudomonas cytochrome c peroxidase. 165 79

Sensitive sandwich enzyme-linked immunosorbent assay (ELISA) systems for the quantification of 3 pathogenic factors of Pseudomonas aeruginosa-alkaline proteinase (aeruginolysin), elastase (pseudolysin ), and exotoxin A-were developed. The maleimide-pyridyl disulfide method was applied for the labeling of rabbit anti-each antigen IgG with horseradish peroxidase (HRP) and the conjugates were used as secondary antibodies (detection antibodies) in the ELISA systems. The EDTA, a chelating agent, was added to the buffers for sample and detection antibody, which inhibited the degradation of IgG by elastase derived from P. aeruginosa for improving the assay precision. The ELISA systems using the HRP-labeled detection antibodies produced by the maleimide-pyridyl disulfide method exhibited higher sensitivity than previously reported methods. The detection limits for alkaline proteinase, elastase, and exotoxin A were 18 pg/ml, 34 pg/ml, and 22 pg/ml, respectively. The intra-assay coefficients of variation for alkaline proteinase, elastase, and exotoxin A were 3.4%-5.0%, 1.9%-3.5%, and 1.3%-5.4%, respectively. These ELISA systems exhibited good inter-assay precision, non-cross-reactivity, dilution linearity, and recovery . Employing these ELISA systems, we revealed that pathogenic factor concentrations were different among the P. aeruginosa strains tested, which may relate to the different pathogenicity of each strain.
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PMID:Reliable enzyme-linked immunosorbent assay systems for pathogenic factors of Pseudomonas aeruginosa alkaline proteinase, elastase, and exotoxin A: a comparison of methods for labeling detection antibodies with horseradish peroxidase. 1809 33