Nicotinamide Adenine Dinucleotide Analogues Research Articles

Purification and properties of cytoplasmic and mitochondrial malate dehydrogenases of Physarum polycephalum.

Two isoenzymes of malate dehydrogenase (MDH) were demonstrated in plasmodia of Physarum polycephalum by polyacrylamide-gel electrophoresis. The more "cathodal" form was uniquely associated with mitochondria (M-MDH) and the other form was found in the soluble cytoplasm (S-MDH). The isoenzymes were separated by acetone fractionation of soluble plasmodial homogenates acidified to pH 5.0. The M-MDH was purified 201-fold by cetylpyridinium chloride treatment, fractionation with ammonium sulfate, gradient elution from sulfoethyl cellulose at pH 6.0, and Sephadex G-100 chromatography. The S-MDH was purified 155-fold by ammonium sulfate fractionation, diethylaminoethyl cellulose chromatography, gradient elution from sulfoethyl cellulose at pH 5.5, and Sephadex G-100 chromatography. The optimal cis-oxalacetate concentrations were 0.35 mM for M-MDH and 0.25 mM for S-MDH, and the optimal pH for both isoenzymes was 7.6 for oxalacetate reduction. The optimal l-malate concentrations were 5 mM for S-MDH and 6 mM for M-MDH, and both isoenzymes exhibited an optimal pH of 10.0 for L-malate oxidation. The Michaelis constants of S-MDH and M-MDH served to discriminate between the isoenzymes. The S-MDH was more heat-stable than the M-MDH. High concentrations of oxalacetate and malate inhibited S-MDH more than M-MDH. The isoenzymes were further distinguished by their utilization of analogues of nicotinamide adenine dinucleotide. Many properties of the Physarum isoenzymes were similar to those of more complex organisms, especially vertebrates.

Identification of Supernatant and Mitochondrial Isozymes of Malate Dehydrogenase on Electropherograms Applied to the Taxonomic Discrimination of Walleye (Stizostedion vitreum vitreum), Sauger (S. canadense), and Suspected Interspecific Hybrid Fishes

Walleye (Stizostedion vitreum vitreum) and sauger (S. canadense) commonly yield four isozymes of malate dehydrogenase (MDH) in electropherograms of white muscle extracts from individual homozygous fish. The heat stability of each isozyme as well as reactivity with nicotinamide-adenine dinucleotide (NAD) and NAD analogues have been investigated by measuring the density of each isozyme directly on the electropherograms, after the usual specific staining procedure that links malate oxidation to tetrazolium dye reduction. At least two kinds of supernatant and one kind of mitochondrial MDH in individual fish of each species was demonstrated.Relative abundance of the various isozymes varies with the tissue of origin. In liver, only one heat stable isozyme is observed in both species, and it is likely constituted mainly of supernatant MDH. In heart tissue four isozymes are observed and the liver supernatant form also predominates. In white muscle the two kinds of supernatant MDH appear to be synthesized in comparable amounts and, including the mitochondrial MDH, four isozymes of nearly equivalent catalytic activity are produced. In walleye the three kinds of MDH homozygotes all showed similar enzymatic properties of their corresponding isozymes.In contrast to the polymorphic nature of walleye MDH isozymes the closely related sauger are monomorphic for MDH, and a number of analogous isozymes have identical electrophoretic mobility in the two species. More significantly, the major mitochondrial MDH isozymes have different mobility in each species. This fact is suggested as a taxonomic criterion to distinguish the two species. Some very rare fishes, evidently interspecific hybrids, produced three mitochondrial MDH isozymes. It was also possible to hybridize walleye and sauger MDH isozymes in vitro to produce phenotypes indistinguishable from presumed wild hybrid fishes.

A fluorescent analog of nicotinamide adenine dinucleotide.

Nicotinamide 1,N(6)-ethenoadenine dinucleotide, a fluorescent analog of the coenzyme nicotinamide adenine dinucleotide, has been synthesized by the reaction of chloroacetaldehyde with the coenzyme. The technical fluorescence emission maximum of the analog is 410 nm, upon excitation at 300 nm. Its fluorescence yield is about 8% of that of the 1,N(6)-ethenoadenine 5'-phosphate, and its fluorescence lifteime is shorter. Upon hydrolysis of the modified coenzyme analog with Neurospora crassa NADase or phosphodiesterase I at room temperature, the intensity of fluorescence was increased 10-fold, corresponding to separation of the nicotinamide and ethenoadenine rings. The spectroscopic results with nicotinamide 1,N(6)-ethenoadenine dinucleotide are consistent with the concept of an intramolecular interaction between the modified adenine and pyridine moieties of the dinucleotide that is disrupted by enzymatic hydrolysis. The fluorescent analog showed reasonable activity as a substitute for NAD(+) in four different dehydrogenase-catalyzed reactions.

Carbohydrate Metabolism During Morphogenesis of Coprinus lagopus ( sensu Buller)

The occurrence and properties of enzymes of carbohydrate metabolism were studied during dikaryotic fruiting of the mushroom Coprinus lagopus. Enzymes of hexose monophosphate catabolism, sugar alcohol (polyol) dehydrogenases (DH), and trehalase occurred throughout development. The ratio of xylitol DH to sorbitol DH was greater than unity in both monokaryotic mycelium and dikaryotic fruit body caps, whereas this ratio decreased in the stipe (stalk) tissue. Xylitol DH and sorbitol DH were both dependent upon nicotinamide adenine dinucleotide (NAD) and showed maximal activity at pH 9. Two separate enzymes were suspected on the basis of preferential utilization of the NAD analogue, thionicotinamide-NAD, by xylitol DH, and this feature was consistent throughout development. An appraisal of the carbohydrate pool revealed trehalose and glucose, with the former predominant in the stipe and the latter in excess in the cap of dikaryotic fruit bodies. Trehalase activity in dialyzed enzyme extracts showed pH optima at acid and alkaline pH levels in monokaryotic mycelium, dikaryotic stipes, and cap tissues.

Interaction of a spin-labeled analog of nicotinamide-adenine dinucleotide with alcohol dehydrogenase. I. Synthesis, kinetics, and electron paramagnetic resonance studies.

ADVERTISEMENT RETURN TO ISSUEPREVArticleNEXTInteraction of a spin-labeled analog of nicotinamide adenine dinucleotide with alcohol dehydrogenase. I. Synthesis, kinetics, and electron paramagnetic resonance studiesHenry WeinerCite this: Biochemistry 1969, 8, 2, 526–533Publication Date (Print):February 1, 1969Publication History Published online1 May 2002Published inissue 1 February 1969https://doi.org/10.1021/bi00830a011RIGHTS & PERMISSIONSArticle Views86Altmetric-Citations77LEARN ABOUT THESE METRICSArticle Views are the COUNTER-compliant sum of full text article downloads since November 2008 (both PDF and HTML) across all institutions and individuals. These metrics are regularly updated to reflect usage leading up to the last few days.Citations are the number of other articles citing this article, calculated by Crossref and updated daily. Find more information about Crossref citation counts.The Altmetric Attention Score is a quantitative measure of the attention that a research article has received online. Clicking on the donut icon will load a page at altmetric.com with additional details about the score and the social media presence for the given article. Find more information on the Altmetric Attention Score and how the score is calculated. Share Add toView InAdd Full Text with ReferenceAdd Description ExportRISCitationCitation and abstractCitation and referencesMore Options Share onFacebookTwitterWechatLinked InReddit PDF (775 KB) Get e-Alerts Get e-Alerts

Interaction of a spin-labeled analog of nicotinamide-adenine dinucleotide with alcohol dehydrogenase. II. Proton relaxation rate and electron paramagnetic resonance studies of binary and ternary complexes.

Interaction of a spin-labeled analog of nicotinamide-adenine dinucleotide with alcohol dehydrogenase. II. Proton relaxation rate and electron paramagnetic resonance studies of binary and ternary complexes.

Identification of Analogues of Nicotinamide Adenine Dinucleotide among the Metabolites of 2,6-Diaminopurine in Mammalian Cells

Experimental evidence indicates that an analogue of nicotinamide adenine dinucleotide in which the adenine moiety is replaced by 2,6-diaminopurine is formed when cultured mammalian cells are grown in the presence of 2,6-diaminopurine. It is also likely that the corresponding analogue containing nicotinic acid is formed when both 2,6-diaminopurine and nicotinic acid are added to the culture medium.

The preparation of some 4-substituted nicotinic acids and nicotinamides.

Nicotinamide adenine dinucleotide (NAD) analogues in which the 4-position of the pyridine ring carries a substituent are potential inhibitors of the glycolytic process by which many cancer cells derive an appreciable proportion of their energy requirements. Such analogues should be produced in vivo by the administration of 4-substituted nicotinic acid derivatives.The preparation of the following derivatives is now described: 4-hydroxy-, 4-methoxy-, 4-butoxy-, 4-(2-hydroxyethylthio)-, 4-S-cysteinyl-, 4-amino-, 4-anilino-, and 4-benzylamino-nicotinic acid and -nicotinamide. 4-Ethoxy- and 4-(2-bromoethylthio)-nicotinic acid and 4-benzylthionicotinamide were also obtained.A preliminary report on the biological activity of some of the derivatives is given.

AN UNUSUAL ISOZYME OF LACTATE DEHYDROGENASE IN MATURE TESTES: LOCALIZATION, ONTOGENY, AND KINETIC PROPERTIES.

SummaryOne or more electrophoretically unusual isozymes of LDH have been demonstrated in homogenates of mature testes from some species. Analyses of the isozymic composition of many other tissues, developing testes, and seminal fluid revealed that these unusual forms, or “band X” isozymes, are present only in postpubertal testes and are localized in the differentiating germ cells of the seminiferous tubules. Rates of reactivity with certain alpha‐hydroxy acids and analogs of nicotinamide adenine dinucleotide differentiated some of the “band X” isozymes from the usual LDH isozymes. Results of dissociation and recombination studies on homogenates of human and rabbit testes indicated a similarity in structure between “band X” and the other isozymes.The ontogeny and characteristic distribution of the “band X” isozymes suggested that each fulfills a specific metabolic function. As yet the nature of this metabolic assignment has not been determined.

NICOTINIC ACID ANALOGS: EFFECTS ON RESPONSE OF CHICK EMBRYOS AND HENS TO ORGANOPHOSPHATE TOXICANTS.

Embryonic abnormalities are known to be induced in chick embryos when they are injected with certain organophosphate insecticides prior to incubation. The marked teratogenic effects of Bidrin [3-(dimethoxyphosphinyloxy) N, N-dimethyl-cis-crotonamide] can be alleviated by many analogs of nicotinamide and nicotinamide adenine dinucleotide. Successive hydroxylation and N-dealkylation of Bidrin in the egg are not greatly altered by injection of nicotinamide. The delayed neurotoxicity in adult hens after administering tri-o-cresyl phosphate is also partially relieved by administering certain nicotinamide analogs.

MOLECULAR HETEROGENEITY OF ENZYMES: MALATE DEHYDROGENASE OF EUGLENA.

Experiments with analogs of the coenzyme nicotinamide adenine dinucleotide demonstrate that the molecular forms of malate dehydrogenase in Euglena vary with the nutritional environment. Electrophoretic separations on starch gel show that Euglena grown on autotrophic medium has a malate dehydrogenase which is lacking in Euglena grown on heterotrophic medium.

SYNTHESIS OF NICOTINAMIDE ADENINEDINUCLEOTIDE ANALOGS AND THEIR COENZYMATIC ACTIVITIES.

Five NAD analogs have been synthesized in which the adenosine portion of NAD was replaced by deoxyadenosine, guanosine, cytidine, uridine and thymidine, and their comparative coenzymatic activity with that of NAD was investigated with yeast alcohol dehydrogenase, beef liver glucose dehydrogenase and rabbit muscle lactic dehydrogenase. The influence of the analogs on the reaction rate of NAD was also studied.