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Query: EC:1.8.1.4 (
diaphorase
)
2,754
document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)
Previous papers in the series have shown that the surface membranes of herpesvirus-infected cells acquire new immunological specificities and that purified infected cell membrane preparations, characterized by their physical properties rather than topology in the cell, contain new glycoproteins genetically determined by the virus. In this study, we prepared purified plasma membrane identified by its 5' nucleotidase, fucose, and reduced
nicotinamide
adenine dinucleotide-
diaphorase
content. Analysis of the membrane proteins and glycoproteins by electrophoresis in acrylamide gels indicated the following. (i) Purified plasma membranes from infected cells contained two sets of proteins, i.e., host proteins were present both before and after infection and viral proteins were present only after infection. (ii) After infection, no appreciable selective or nonselective loss of host proteins from membranes was demonstrable. However, no new host proteins were made. (iii) Electropherograms of plasma membrane proteins from infected cells indicated the presence of at least 12 virus-specific proteins ranging in molecular weight from 25 x 10(3) to 126 x 10(3) daltons. Of these, at least nine were glycosylated. Proteins and glycoproteins with similar electrophoretic mobilities but in somewhat different ratios were also present in preparations of highly purified virions.
...
PMID:Proteins specified by herpes simplex virus. VI. Viral proteins in the plasma membrane. 411 36
Total reduced
nicotinamide
adenine dinucleotide (NADH) and reduced
nicotinamide
adenine dinucleotide phosphate (NADPH)
diaphorase
activities were examined in human neutrophils. Approximately two-thirds of each enzyme activity was located in the granule fraction with the remainder in the soluble. The activities in a 27,000 x g supernatant from a sonic extract of human polymorphonuclear leukocytes were characterized. Both NADH and NADPH diaphorase were insensitive to cyanide and azide and showed greater activity at acid pH. K(m) values for nitroblue tetrazolium were not markedly different (33 muM with NADH and 12 muM with NADPH), but there was a 40-fold difference in K(m) for the reduced pyridine nucleotides (10 muM with NADH and 400 muM for NADPH). Since the intracellular concentration of both nucleotides is estimated to be about 50 muM, it is much more likely, from a kinetic argument, that the respiratory burst of phagocytosis is intiated by the oxidation of NADH rather than of NADPH.
...
PMID:Reduced nicotinamide adenine dinucleotide and reduced nicotinamide adenine dinucleotide phosphate diaphorase activity in human polymorphonuclear leukocytes. 415 6
The cytochemical localization, by conventional methods, of lactate and glyceraldehyde-3-phosphate dehydrogenases is limited, firstly, by the solubility of these enzymes in aqueous media and, secondly, by the dependence of the final electron flow from reduced
nicotinamide
-adenine dinucleotide (NADH) to the tetrazolium on tissue
diaphorase
activity: localization is therefore that of the
diaphorase
, which in rabbit adductor magnus is mitochondrial. NADH has been found to have great affinity to bind in the sarcoplasmic reticulum, and, therefore, if it is generated freely in the incubation media containing 2,2',5,5'-tetra-p-nitrophenyl-3,3'-(3,3'-dimethoxy-4,4'-phenylene)-ditetrazolium chloride (TNBT) and N-methyl phenazonium methyl sulfate (PMS), it can bind there and cause a false staining. Since such a production of NADH can readily occur in the incubation media for glycolytic dehydrogenases due to diffusion of these soluble enzymes from tissue sections, the prevention of enzyme solubilization is extremely important. Fixation in formaldehyde prevented such enzyme diffusion, while at the same time sufficient activity persisted to allow for adequate staining. The incubation media contained PMS, so that the staining system was largely independent of tissue
diaphorase
activity. Application of these methods to adductor magnus of rabbit revealed by light microscopy, for both enzymes, a fine network which was shown by electron microscopy to represent staining of the sarcoplasmic reticulum. Mitochondria also reacted. These findings add further support for the notion that the sarcoplasmic reticulum is probably involved in glycolytic activity.
...
PMID:Cytochemical localization of two glycolytic dehydrogenases in white skeletal muscle. 428 29
Optochin-resistant mutant and wild-type diaphorases were purified approximately 300-fold by a combination of batch adsorption and column chromatography with diethylaminoethyl cellulose, and were characterized with regard to their pH optima, sensitivity to optochin inhibition and heat inactivation, Michaelis constants with flavine mononucleotide (FMN) and reduced
nicotinamide
adenine dinucleotide (NADH), and inhibition constants with optochin hydrochloride. The pH optima of the purified diaphorases were similar, but the purified diaphorases from the optochin-resistant strains were approximately four to five times more resistant to heat inactivation at 45 C than was the wild-type
diaphorase
. Purified
diaphorase
preparations from the optochin-resistant pneumococci had greater activities per milligram of protein and were more resistant to optochin inhibition than the preparation from the optochin-sensitive pneumococcus. Michaelis constants for FMN and NADH were similar; however, the inhibition constants of the optochin-resistant diaphorases were four to eight times greater than that of the optochin-sensitive
diaphorase
. Optochin hydrochloride produced a noncompetitive type of inhibition with FMN as substrate but a competitive type of inhibition with NADH as substrate. Optochin hydrochloride produced an approximately 10-fold increase in the Michaelis constant for NADH. The concentration of drug required to produce this effect was, however, greater with the mutant diaphorases than with the wild-type
diaphorase
. Optochin hydrochloride quenched the fluorescence of riboflavine. This phenomenon did not appear to be related to the
diaphorase
-inhibitory activity of the drug, however, since the pH requirements of the two reactions were different. Quenching of riboflavine fluorescence by optochin hydrochloride increased with a rise in pH, whereas inhibition of
diaphorase
activity by optochin hydrochloride was greater at pH 6.8 than at pH 7.6.
...
PMID:Purification and properties of mutant and wild-type diaphorases from Diplococcus pneumoniae. 438 90
Evidence that the bactericidal ability and the stimulated oxidative metabolism of leukocytes appear in parallel during fetal development of the Minnesota Miniature pig has been obtained by application of the techniques applied to studies of human cells. It was demonstrated that leukocytes from 87- to 90-day fetuses were fully capable of ingesting Staphylococcus aureus but greatly diminished in bactericidal capacity as compared to leukocytes of older fetuses and adults. Although resting levels of oxygen consumption and hexose monophosphate pathway activity of leukocytes from the younger fetuses compared well with those of leukocytes from older animals, the phagocytosis-stimulated increments of metabolism were much less at 87 to 90 days of gestation than at later developmental stages. Both bactericidal capacity and increased metabolism of leukocytes reach adult levels by 100 days of gestation (normal gestation period of 115 to 120 days). Acrylamide gels stained for reduced
nicotinamide
adenine dinucleotide (NADH) and NADH phosphate (NADPH)
diaphorase
activity after disc electrophoresis of leukocyte extracts revealed normal mobility and intensity of NADH diaphorase bands. Three NADPH diaphorase bands were present in adult leukocyte extracts. Only the fast-migrating NADPH diaphorase band of 87- to 90-day cells stained with decreased intensity. This "deficiency" was no longer present at the later fetal period. The fast-migrating NADPH diaphorase band may represent an electron transfer protein which functions in cyanide-insensitive respiration of the leukocytes of the pig.
...
PMID:Development of bactericidal capacity and phagocytosis-associated metabolism of fetal pig leukocytes. 463
alpha-Glycerophosphate oxidase, in a strain of Streptococcus faecium, was found to be adaptive to aerated conditions of growth. The enzyme was purified and found to mediate electron transfer from alpha-glycerophosphate to O(2), with the production of stoichiometric concentrations of H(2)O(2) and dihydroxyacetone phosphate. The enzyme is an anionic flavoprotein, with flavine adenine dinucleotide as the apparent prosthetic group. By manometric methods, a K(m) of 6 x 10(-3)m, with reference to substrate concentration, was obtained. An active reduced
nicotinamide
adenine dinucleotide
diaphorase
was closely associated with this enzyme in chromatographic mobility on ECTEOLA-cellulose. The purified alpha-glycerophosphate oxidase was not inhibited by KCN, azide, or sulfhydryl reagents, nor was it stimulated by alpha-lipoate, yeast extract, or other supplements.
...
PMID:Alpha-glycerophosphate oxidase in Streptococcus faecium F 24. 578 98
Striatal neurons containing both somatostatin- and avian pancreatic polypeptide (APP)-like immunoreactivities have been shown to be selectively stained by the histochemical method for
nicotinamide
adenine dinucleotide phosphate (NADPH)-
diaphorase
activity. In the present study, we have utilized this histochemical technique to examine the morphology of these striatal neurons at the light and electron microscopic levels. Our results indicate that the striatal somatostatin/APP/NADPH-diaphorase neurons occur throughout the striatum and have long, aspiny dendrites, oval, invaginated nuclei with prominent nucleoli, and receive few axosomatic contacts. These cells appear to correspond to a population of medium-sized aspiny interneurons reported previously in Golgi and electron microscopic studies of the striatum.
...
PMID:Striatal neurons containing both somatostatin- and avian pancreatic polypeptide (APP)-like immunoreactivities and NADPH-diaphorase activity: a light and electron microscopic study. 613 32
A single administration of chlorophos (trichlorophon) solution (600 mg/kg) (LD50) results in vacuolar distrophy appearing in the white rat liver and is especially pronounced in 2-4 days. Thirty minutes after the poisonous chemical is administered, butyrilcholinesterase (BChE) activity is inhibited by 90%, somewhat later oxidation-reduction enzymes activity decreases and alkaline phosphatase (APh) activity increases. Cytoplasm of hepatocytes is filled with glycogene and nearly deprived of RNA. Owing to the cytophotometric analysis of the enzymatic activity and the stereologic morphometry method, it has been possible to reveal a certain synchronism in the development of distrophic processes, in a decreasing activity of the oxidation-reduction enzymes and in a disturbed synthesis of glycogene and RNA. On the 6th day after chlorophos has been administered, succinate dehydrogenase and
nicotinamide
-adenine-dinucleotide-phosphate-
diaphorase
activity, as well as contents and distribution of RNA in hepatocytes reach their control values. BChE and APh activity does not restore. During the whole experiment there is not any statistically significant change in the volumetric part of the sinusoid capillaries and in the stellate reticuloendotheliocytes. Thus, the main effect of chlorophos action is a specific inhibition of ChE, that results in certain structural changes and in changes of the histoenzymatic parameters of the liver.
...
PMID:[Morpho-functional changes in the liver after exposure to cholinesterase inhibitors]. 619 75
Experiments were performed to determine whether conditions which cause the rapid loss of nitrate reductase activity in Neurospora crassa mycelia were accompanied by the loss of antigenically detectable nitrate reductase protein. When mycelia with nitrate reductase activity were transferred to ammonia media, there was a rapid loss in the reduced
nicotinamide
adenine dinucleotide-nitrate reductase activity plus the parallel loss of the reduced
nicotinamide
adenine dinucleotide-
diaphorase
and the reduced methyl viologen-nitrate reductase activities associated with the nitrate reductase. In addition, there was the loss of cross-reacting material to anti-nitrate reductase antisera that was concomitant with the loss of nitrate reductase activity. When mycelia were exposed to either ammonia plus cycloheximide, nitrate plus cycloheximide, or nitrogen-free media, or to media which lacked an assimilable carbon source, the amount of cross-reacting material declined in concert with the nitrate reductase activity. The mutant nit-6, which lacks nitrite reductase activity, was exposed to ammonia or nitrate plus cycloheximide media. The nitrate reductase and the amount of cross-reacting material declined together as in the wild-type mycelia. We conclude that the loss of nitrate reductase activity was accompanied by the specific loss of this protein and that no pool of inactivated nitrate reductase molecules existed.
...
PMID:Repression of nitrate reductase activity and loss of antigenically detectable protein in Neurospora crassa. 644 48
A comparison was made of muscle from two locations in both the longissimus and the semitendinous muscles of normal and malignant hyperthermia-susceptible swine. Serial frozen sections were stained for alkali-stable adenosine triphosphatase (ATPase), phosphorylase, and the oxidative enzymes succinate dehydrogenase and reduced
nicotinamide
adenine dinucleotide (NADH)-
diaphorase
. Myofiber types were identified on the basis of these staining reactions. There was no consistent statistically significant difference between muscle from normal and muscle from susceptible swine with any system of fiber classification. This is contrary to several published reports but consistent with physiologic studies which indicate that both oxidative and glycolytic pathways are abnormally active during the onset of malignant hyperthermia.
...
PMID:Histochemical observations on muscle from normal and malignant hyperthermia-susceptible swine. 644 66
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