Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:1.17.3.2 (xanthine oxidase)
 8,383 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

A blotting method is described to detect enzymes that do not normally yield a colored product. The method can be used for dot blotting as well as blotting after gel electrophoresis of many enzymes if the reactions they catalyze can be coupled to an oxidase or a dehydrogenase. The latter, designated "auxiliary enzymes," are preimmobilized on membranes of nitrocellulose or positively charged nylon and the reaction they catalyze is coupled with reduction of tetrazolium salt to yield colored formazan on areas of the transfer membrane occupied by the blotted enzymes. In the examples reported here, preimmobilized glucose oxidase, L-amino acid oxidase, xanthine oxidase, malate dehydrogenase, and a mixture of hexokinase and glucose-6-phosphate dehydrogenase were used as auxiliary enzymes to detect blotted invertase, leucine aminopeptidase, purine nucleoside phosphorylase, fumarase, and adenylate kinase, respectively. Detection limits varied, but never exceeded 100 ng for these enzymes. After blotting from polyacrylamide gels, the fumarase assay was the most sensitive of those investigated, detecting 10 ng of enzyme used for electrophoresis. Invertase, a glycoprotein, was detected with higher sensitivity on nitrocellulose membranes when concanavalin A was present on the membrane in addition to the auxiliary enzyme, glucose oxidase. On blots from isoelectric focusing gels, the assay detected two isozymes of purine nucleoside phosphorylase in a sample from calf spleen and at least five isozymes of this enzyme in lysates from human red cells.
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PMID:Activity staining of blotted enzymes by reaction coupling with transfer membrane-immobilized auxiliary enzymes. 245 38

The freshwater murrel, Channa punctatus, was exposed to a sublethal concentration of mercuric chloride (3 micrograms/liter) for 120 days and the following effects were examined: changes in the levels of glucose and lactic acid in blood and of glycogen and lactic acid in liver and muscles; rate of absorption of glucose from the intestine; and changes in the activities of glucose-6-phosphatase (G-6-Pase), hexokinase, lactate dehydrogenase (LDH), pyruvate dehydrogenase (PDH), succinate dehydrogenase (SDH), malate dehydrogenase (MDH), glutamate dehydrogenase (GDH), L-amino acid oxidase (AO), and xanthine oxidase (XO) in brain, gills, intestine, kidney, liver, and muscles. Mercury-treated fish were hypoglycemic and hypolactemic. The glycogen content of liver and muscles remained unaltered but the muscle lactic acid level decreased significantly. The rate of intestinal absorption of glucose was reduced significantly by exposure to mercury. G-6-Pase activity was decreased in all the tissues. Hexokinase activity also decreased in mercury-exposed fish but it was significant only in intestine, kidney, and liver. The activities of LDH, PDH, SDH, and MDH also were decreased significantly except LDH in brain and MDH in kidney where an insignificant decrease and an insignificant increase, respectively, were recorded. GDH and AO activities were elevated in most of the tissues except GDH in gills, and AO in gills and muscles where a decrease was observed. XO activity in brain, gills, and kidneys was significantly elevated, but no marked alteration was noted in other tissues.
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PMID:Effect of mercuric chloride on some biochemical and physiological parameters of the freshwater murrel, Channa punctatus. 608 7

The chain oxidation of glyceraldehyde-3-phosphate dehydrogenase.NADH by perhydroxyl radicals and propagated by molecular oxygen was studied by the xanthine-xanthine oxidase system, 60Co gamma-ray, and pulse radiolysis. The chain length, amount of NADH oxidized per HO2 generated, increases with increasing acidity of the medium and reaches a value of 73 at pH 5.0. The rate constant for the oxidation of the glyceraldehyde-3-phosphate dehydrogenase.NADH complex by HO2 was estimated to be 2 X 10(7) M-1 S-1 at ambient temperatures (23-24 degrees C). Rate studies as a function of pH indicate that O2- is unreactive toward the glyceraldehyde-3-phosphate dehydrogenase.NADH complex. Other dehydrogenases (malate dehydrogenase, glutamate dehydrogenase, and isocitric dehydrogenase) studied showed no catalytic activity in the oxidation of NADH by HO2/O2-.
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PMID:Glyceraldehyde-3-phosphate dehydrogenase-catalyzed chain oxidation of reduced nicotinamide adenine dinucleotide by perhydroxyl radicals. 718 97

Two types of superoxide dismutase (SOD) have been found in Brucella abortus, a cytosolic Mn-SOD and a Cu/Zn-SOD of unknown location. We sought to determine the subcellular location of Cu/Zn-SOD in B. abortus ST 19. We report a modified spheroplasting procedure for the release of periplasmic contents from B. abortus cells using a dipolar ionic detergent, Zwittergent 316. This detergent, used in place of EDTA, destabilizes the outer membrane sufficiently to allow penetration of lysozyme and the subsequent selective release of periplasmic proteins by osmotic shock. Cytoplasmic cross-contamination of periplasmic fractions was assessed by assaying for malate dehydrogenase activity. Cyanide-sensitive and cyanide-insensitive SOD activity was measured by both the xanthine oxidase-cytochrome c method and a hematoxylin assay. Results suggest that B. abortus Cu/Zn-SOD activity is periplasmic. This zwittergent-lysozyme extraction procedure may be applicable to the separation, isolation and characterization of many other periplasmic proteins of B. abortus and other Gram-negative organisms especially when cytosolic contamination is undesirable.
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PMID:Periplasmic location of Brucella abortus Cu/Zn superoxide dismutase. 816 Mar 46