Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UMLS:C0272170 (SDS)
 50,377 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Macrophage deactivation factor (MDF) in P815 tumor cell-conditioned medium was assayed by its suppression of the ability of activated mouse peritoneal macrophages to release hydrogen peroxide. MDF displayed properties of a soluble protein(s) associated with both low (8-25,000) and high (greater than 450,000) Mr fractions. MDF was purified 6,140-fold by a seven-step procedure: extraction with acid-ethanol; precipitation with ether; and fractionation on gel filtration, anion-exchange, diphenyl reversed-phase and C4 reversed-phase HPLC columns, the last column twice. The final preparation contained two species: (a) a approximately 13,000 Mr band on reducing or nonreducing SDS-PAGE and on autoradiograms after radioiodination with chloramine T, and (b) a 66,000 Mr species ranging from approximately 5% to approximately 50% of the protein detectable by silver strain. The 66,000 Mr species was identified as albumin from its NH2-terminal amino acid sequence. However, no amino acid sequence could be obtained for the approximately 13,000 Mr species, either in fluid phase or after electroelution of the corresponding SDS-PAGE band. Thus, approximately 13,000 Mr MDF associates tightly with albumin through a variety of separation techniques, and may have a blocked NH2 terminus. Purified MDF afforded 50% inhibition of activated macrophage H2O2 releasing capacity at a concentration of 1-10 nM. Separation of MDF from most higher Mr moieties was associated with disproportionately small increases in specific activity, suggesting MDF might be partially inactivated by purification. As purified, MDF was approximately 1,000-fold less potent at deactivating macrophages than TGF-beta. Antibodies that neutralized the macrophage-deactivating effect of TGF-beta did not inhibit deactivation by MDF.
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PMID:Purification of macrophage deactivating factor. 210 38

Purified recombinant human monocyte plasminogen activator inhibitor 2 (PAI-2) retained inhibitory activity after exposure to a number of oxidants, including hypochlorite anion (OCl-), chloramine-T (CT) and hydrogen peroxide (H2O2). Analysis of PAI-2 exposed to oxidants by gel filtration chromatography and SDS-PAGE indicated that although the protein could no longer be detected by silver staining, this was not due to fragmentation of the PAI-2 molecule. The sensitivity of a number of serine protease inhibitors (serpins), (eg. alpha 1 proteinase inhibitor (alpha 1PI) and plasminogen activator inhibitor 1 (PAI-1] to oxidative inactivation has been attributed to oxidation of reactive site methionine residues and/or tertiary structural modifications. The relevance of these phenomena and the potential for PAI-2 to be used as a therapeutic inhibitor of urokinase (uPA)-dependent proteolysis during inflammation and tumour metastasis is discussed.
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PMID:Plasminogen activator inhibitor 2 (PAI-2) is not inactivated by exposure to oxidants which can be released from activated neutrophils. 215 27

Incubation of a 90-kilodalton ferritin repressor protein (FRP), either free or complexed with an L-ferritin transcript, with hemin or Co3+-protoporphyrin IX prevented subsequent repression of ferritin synthesis in a wheat germ extract. Neither FeCl3 in combinations with H2O2, nor Fe3+ or Fe2+ chelated with EDTA, nor Zn2+-protoporphyrin IX, nor protoporphyrin IX caused significant inactivation of FRP. FRP that had been inactivated by hemin remained chemically intact, as revealed by SDS-polyacrylamide gel electrophoresis. Inclusion of chelators of iron or free radical scavengers did not alter the inactivation produced by hemin. These and other results indicate that hemin derepresses ferritin synthesis in vitro.
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PMID:Derepression of ferritin messenger RNA translation by hemin in vitro. 229 94

A glutathione peroxidase was purified from bovine ciliary body by ammonium sulfate fractionation. Sephacryl S-300 gel filtration, diethylaminoethyl (DEAE)-cellulose chromatography and hydroxyapatite chromatography. The purified enzyme has an apparent mw of 112 kDa by gel filtration and 29 kDa by SDS-polyacrylamide gel electrophoresis. The enzyme therefore is composed of four identical subunits. The ciliary enzyme is active with H2O2 (25), cumene hydroperoxide (170), t-butyl hydroperoxide (22), triphenylcarbinyl hydroperoxide (12), linoleic hydroperoxide (34) and 5-phenylpentenyl hydroperoxide (22): the numbers after substrates are K'm in microM. Glutathione is essential for the reaction; L-cysteine, dithiothreitol and 2-mercaptoethanol are inactive. Mercaptosuccinate (10 microM) inhibits the enzyme competitively (Ki = 7 microM) when cumene hydroperoxide is substrate, and uncompetitively (Ki = 10 microM) when H2O2 is substrate. No selenium was found in the enzyme by the fluorometric assay with 2.3-diaminonaphthalene. The enzyme demonstrates no glutathione S-transferase activity when tested with 1-chloro-2,4-dinitrobenzene, and several other compounds. A partial sequence of the enzyme shows some similarities both to Se-glutathione peroxidases and a glutathione S-transferase isozyme.
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PMID:Non-selenium glutathione peroxidase without glutathione S-transferase activity from bovine ciliary body. 237 54

Lipid peroxide and SOD were selected as free radical related substances and system for their elimination, and detection was evaluated. NADPH-Cytochrome c reductase-Neotetrazolium (NT) method (Mic-NT method) and Xanthine oxidase-Nitrotetrazolium Blue method (XOD-NTB method) are current detection methods of SOD activities. They are based on the O2-specific reaction. Minimum detectable amount of SOD by the Mic-NT method and XOD-NTB method was about 15 ng and 200 ng, respectively. On the other hand, an XOD-NH2OH method which detects SOD activities based on the O2-specific oxidation reaction showed the minimum detectable amount of 2.5 ng. Consequently, SOD-detecting sensitivity of these methods was found to be in the following order: XOD-NH2OH method greater than Mic-NT method greater than XOD-NTB method. In addition, albumin caused a positive error in all three methods. With a monoclonal antibody-aided SOD-analyzing method (EIA method), the minimum detectable amount of SOD was 0.2 ng. The isoenzymes of SOD (Cu, Zn-SOD and Mn-SOD) could be detected separately by 1. deactivating Cu, Zn-SOD with CN- or H2O2 and regarding the remaining activity as Mn-SOD and 2. by deactivating Mn-SOD selectively through pretreatment of the sample with SDS and regarding the remaining activity as Cu, Zn-SOD. TBA method (Yagi's method) has been used frequently for the measurement of serum lipid peroxide.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:[Detection methods of free radical related substances and the system for their elimination]. 260 53

The extrinsic 33-kDa protein (P33) was cross-linked covalently to the binding site on P33-depleted PSII preparations which is responsible for reconstitution of photosynthetic water oxidation after PSII preparations have been washed with 1 M CaCl2. Conditions were found in which more than half of the cross-linked protein complexes formed in the PSII preparations retained the ability to catalyze the oxidation of water. The complex is composed of the P33 cross-linked to the D1 and D2 proteins and a 34-kDa protein, which is present in lower abundance than the other three proteins. After solubilization of the membranes with SDS and purification by preparative SDS-PAGE, the complex retains bound manganese and can catalyze the conversion of H2O2 to O2. Calcium and chloride increased the catalase activity of the purified cross-linked complex while lanthanum or hydroxylamine abolished the activity. By use of the specific activity of the H2O2-dependent reaction to follow the extent of purification of the cross-linked complex, the most highly purified complex was determined to contain 0.34 microgram of manganese/180 micrograms of protein. The mole ratio of Mn/protein was calculated to range from 3.6 to 4.5 depending on the assumed stoichiometry of the protein subunits. The results presented here provide direct evidence that one or more of the three proteins that have cross-linked to the P33 are responsible for binding the manganese of the oxygen-evolving complex.
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PMID:Manganese-binding proteins of the oxygen-evolving complex. 267 50

Patulolides are a group of 12-membered macrolide antibiotics produced by Penicillium urticae S11R59. An enzyme involved in the conversion of patulolide C to patulolide A was purified from P. urticae S11R59 and characterized. The enzyme showed a single band on SDS-PAGE and molecular sieve HPLC both of which indicated a Mr of 86,000, indicating that the enzyme is monomeric. However, the enzyme was separated into two bands of very similar pI's (pI 4.2 and 4.3) by isoelectric focusing. Both bands catalyzed the conversion of patulolide C to patulolide A, as demonstrated by activity staining. The two isoenzymes were proved to be oxidases by the simultaneous production of H2O2 during the conversion of patulolide C to patulolide A. The molar ratio for patulolides C, A and H2O2 was determined to be 1:1:1. The optimum pH and temperature were determined to be 7 and 35-40 degrees C, respectively, and the enzymes were stable at pH 6-9 and 4-40 degrees C. The oxidases showed characteristic absorption at 345 and 450 nm, indicating the presence of flavin as coenzyme. Among several analogues of patulolide C tested, the oxidases showed very narrow substrate-specificity; only patulolide C was oxidized to patulolide A. No enzyme activity for the reverse reaction, i.e. from patulolide A to patulolide C, was present in the cell-free extract of P. urticae S11R59. Patulolide C oxidases therefore play a key role in the biosynthesis of patulolides.
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PMID:Oxidases involved in biosynthesis of macrolide antibiotic patulolides from Penicillium urticae S11R59. 272 90

A new flavoenzyme using molecular oxygen to oxidize L-glutamic acid has been purified to homogeneity, as judged by polyacrylamide gel electrophoresis, from the culture medium of Streptomyces endus. Hydrogen peroxide, 2-oxoglutaric acid and ammonia are formed as products. Among 25 amino acids tested including D-glutamic acid, L-glutamine and L-aspartic acid, only L-glutamic acid is converted. The molecular mass of the enzyme was estimated to be about 90 kDa by gel chromatography and 50 kDa by SDS/PAGE. The subunit contains 1 molecule noncovalently bound FAD. The absorption spectrum shows maxima at 273, 355 and 457 nm and the isoelectric point is at pH 6.2. The Km value for L-glutamic acid in air-saturated phosphate pH 7.0 was estimated to be 1.1 mM, the Km for oxygen was calculated to be 1.86 mM at saturating concentration of L-glutamic acid. The enzymic reaction is inhibited by Ag+ and Hg2+ ions. The enzyme described here distinctly differs from two microbial L-glutamate oxidases purified hitherto, with regard to extremely high substrate specificity and to the subunit structure.
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PMID:A novel L-glutamate oxidase from Streptomyces endus. Purification and properties. 273 5

Eighteen white rabbits were subjected to a 30 per cent TBSA full thickness burn. Wound infection was found 9-13 days after injury and became severe a week or so later. ATPase activities, antioxidation ability, the proteins of erythrocyte membranes, and the Na+ contents of erythrocytes and serum were determined. The Ca++-ATPase activity was elevated during the first 17 days postburn, but showed a decline at the time of severe wound infection; the Na+,K+-ATPase activity showed peaks on postburn days 2 and 6, and then fluctuated above the preburn level. The change in Mg++-ATPase activity was similar to Na+,K+-ATPase. The erythrocyte Na+ content was increased, and the level of serum Na+ was decreased up to postburn day 6. Subsequently the erythrocyte Na+ was reduced and the serum Na+ increased up to day 17 postburn. The percentage of erythrocyte haemolysis in H2O2 was increased after the burn and became markedly so during wound infection, indicating that the antioxidation ability of burned rabbit erythrocytes was markedly impaired. During the period of wound infection, Coomassie blue-stained protein bands in SDS-polyacrylamide gel showed some changes in size and proportion in burned rabbits. For example, the second band was wider, the band 2 to band 1 ratio increased, and band 5 was smaller than before injury. These results seem to show that burn injury, especially when associated with sepsis, may affect both the structure and function of biological membranes.
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PMID:Changes in erythrocyte membranes in burned rabbits. 285 50

The ratio between the amount of oligomycin-sensitivity-conferring protein (OSCP) and the amount of the alpha and beta subunits of F1-ATPase in the mitochondria has been determined by a method combining electrophoresis, electrotransfer and immunotitration with monoclonal antibodies. The peptides separated in SDS-polyacrylamide gel electrophoresis were blotted to nitrocellulose sheets by electrotransfer. The nitrocellulose sheets were incubated with 125I-labelled purified monoclonal antibodies specific to various peptides. The 125I-labelled immune complexes were located by immunodecoration using peroxidase-conjugated second antibodies and the blotted peptides were revealed with H2O2 and alpha-naphthol. The amount of immune complex present on the nitrocellulose was determined by counting the radioactivity present on the spots. The amount of peptide blotted is directly proportional to the amount of protein loaded on the electrophoresis. By comparing standard curves made with the isolated proteins to the values obtained in the presence of various amounts of the membrane-protein complex, one can calculate the content of this peptide in the membrane. It was found that the mitochondrial membrane contains 2 mol of OSCP per mol of F1.
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PMID:Stoichiometry of the oligomycin-sensitivity-conferring protein (OSCP) in the mitochondrial F0F1-ATPase determined by an immunoelectrotransfer blot technique. 286 72


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