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1.
The pyridine nucleotide cycle: presence of a nicotinamide mononucleotide-specific amidohydrolase in Propionibacterium shermanii 总被引:2,自引:0,他引:2
A specific nicotinamide mononucleotide amidohydrolase which catalyzes the stoichiometric conversion of NMN to nicotinate mononucleotide and ammonia has been partially purified from an extract of . The reaction has optimum activity at pH 5.6, a Km of 70 μM, and an experimental activation energy of 14.5 Kcal/mole. The enzyme appears to be highly specific for NMN. Neither free nicotinamide nor NAD, NADH, NADP, NADPH compete with NMN. Numerous substances such as isonicotinic acid hydrazide and quinolinic acid are also without effect. It can be stored at ?15° in 12% glycerol, but is somewhat unstable in the absence of this solvent. The enzyme is composed of a heatstable and a heat-sensitive subunit. This enzyme considerably simplifies the pyridine nucleotide cycle, and may, besides this salvage function for NAD, play a role in B12 biosynthesis and in the bacterial DNA ligase reaction. 相似文献
2.
Gwyn T. Williams Sydney Shall Christopher C. Ford 《Biochimica et Biophysica Acta (BBA)/Molecular Cell Research》1983,762(2):272-280
The NAD pools of Xenopus laevis oocytes and early embryos can be radioactively labelled by microinjection of [adenine- 3H]NAD. This technique is used to study the metabolism of NAD in oocytes and during early development. The rate at which NAD is degraded in vivo has been monitored by determining the rate of transfer of adenine residues from the NAD pool into other nucleotides and polynucleotides. In oocytes, NAD turnover is extremely slow, with a half-life of about 400 h. NAD turnover increases dramatically after fertilisation, and the half-life of the compound decreases to 37 h in 5-h-old embryos and to 10 h in 40-h-old embryos. 2 mM 3-aminobenzamide, a specific inhibitor of poly(ADP-ribose) polymerase, reduces the NAD turnover rate by about 20%, whereas 5 mM isonicotinic acid hydrazide, a specific inhibitor of NAD glycohydrolase, produces no significant inhibition. This indicates that a significant fraction of the considerable NAD turnover observed involves poly(ADP-ribose) polymerase. Our results indicate that poly(ADP-ribose) polymerase is active during early development and suggest that this activity may be involved in one or more aspects of the nuclear metabolism of the embryo. 相似文献
3.
The chemical carcinogen hydrazine is a potent stimulator of guanylate cyclase. In the present investigation we found that three chemical carcinogens structurally related to hydrazine, isonicotinic acid hydrazide, hydrazine sulfate, and dimethylhydrazine, decreased guanylate cyclase activity. It is of interest that hydrazine has been shown to increase DNA synthesis whereas isonicotinic acid hydrazide, hydrazine sulfate, and dimethylhydrazine decrease DNA synthesis. The relationship, if any, linking the guanylate cyclase-cyclic GMP system to DNA synthesis and carcinogenesis remains to be explored. 相似文献
4.
N-Methyl-N-nitrosourea was used to induce auxotrophic, scotochromogenic and isonicotinic acid hydrazide resistant mutants inMycobacterium phlei and its effect was compared with that of nitrosoguanidine. Seventeen auxotrophic mutants requiring arnino acids or vitamins and 52 sootochromosgenic mutants with orange colonies were induced. The frequency of isonicotinic acid hydrazide-resistant mutants increased by two orders of magnitude. 相似文献
5.
Effect of some 3- and 4-substituted pyridines on enzymatic hydrolysis of NAD by rabbit heart muscle NAD-glycohydrolase has been studied. It is shown that some 4-substituted derivatives in contrast with 3-substituted ones produce an inhibitory effect on the enzyme activity. A new efficient inhibitor of rabbit heart muscle NAD-glycohydrolase (I50 = 10(-3) M)--N1-(2-lactyl)-N2-(isonicotinoy)hydrazine, inducing uncompetitive inhibition of hydrolysis of NAD is found. The mechanism of the inhibitory effect of N1-(2-lactyl)-N2-(isonicotinoyl)hydrazine was investigated and the rate equation for enzymatic hydrolysis of NAD in the presence of inhibitor is calculated. It is suggested, that the inhibitory effect of N1-(2-lactyl)-N2-(isonicotinoyl)hydrazine is due to the formation of triple inactive complex inhibitor-enzyme-adenosinediphosphateribose. 相似文献
6.
Metabolism of isonicotinic acid and isoniazid bySarcina sp. led to the formation of two metabolites which were characterised as 2-hydroxyisonicotinic acid and citrazinic acid. The
blue pigment formed during fermentation was shown to be derived from the auto-oxidation of citrazinic acid. 2-Oxo-glutarate
accumulated as the major keto acid when isonicotinic acid or isonicotinic acid hydrazide metabolism was inhibited by 1 mM
sodium arsenite. Isonicotinic acid, 2-hydroxy-isonicotinic acid and 2-oxo-glutarate were oxidised by isonicotinic acid hydrazide
or isonicotinic acid-grown cells; citrazinic acid was, however, not oxidised. Isoniazid hydrazine hydrolase, isonicotinic
acid and 2-hydroxyisonicotinic acid hydroxylases were detected in the cell-free extract ofSarcina sp. grown on isonicotinic acid hydrazide or isonicotinic acid.
Communication no. 2427from Central Drug Research Institute, Lucknow. 相似文献
7.
8.
The authors succeeded in demonstrating accumulation of tyrosine and p-coumaric acid in three day-old roots of maize (Zea mays L.) fed with L-phenylalanine and cinnamic acid. Phenylpyruvic acid applied under the same conditions gave rise to phenylalanine, indicating the presence of the corresponding transferase activity. Even simultaneous application of inhibitors of transaminase activity—hydroxylamine and isonicotinic acid hydrazide—did not result in the formation of p-hydroxyphenylpyruvic acid. 相似文献
9.
The authors succeeded in demonstrating accumulation of tyrosine and p-coumaric acid in three day-old roots of maize (Zea mays L.) fed with L-phenylalanine and einnamie acid. Phenylpyruvic acid applied under the same conditions gave rise to phenylalanine, indicating the presence of the corresponding transferase activity. Even simultaneous application of inhibitors of transaminase activity — hydroxylamine and isonicotinic acid hydrazide — did not result in the formation of p-hydroxyphenylpyruvic acid. 相似文献
10.
The purification and properties of peroxidase in Mycobacterium tuberculosis H37Rv and its possible role in the mechanism of action of isonicotinic acid hydrazide.
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Peroxidase from Mycobacterium tuberculosis H37Rv was purified to homogeneity. The homogeneous protein exhibits catalase and Y (Youatt's)-enzyme activities in addition to peroxidase activity. Further confirmation that the three activities are due to a single enzyme was accomplished by other criteria, such as differential thermal inactivation, sensitivity to different inhibitors, and co-purification. The Y enzyme (peroxidase) was separated from NADase (NAD+ glycohydrolase) inhibitor by gel filtration on Sephadex G-200. The molecular weights of peroxidase and NADase inhibitor, as determined by gel filtration, are 240000 and 98000 respectively. The Y enzyme shows two Km values for both isoniazid (isonicotinic acid hydrazide) and NAD at low and high concentrations. Analysis of the data by Hill plots revealed that the enzyme has one binding site at lower substrate concentrations and more than one at higher substrate concentration. The enzyme contains 6g-atoms of iron/mol. Highly purified preparations of peroxidases from different sources catalyse the Y-enzyme reaction, suggesting that the nature of the reaction may be a peroxidatic oxidation of isoniazid. Moreover, the Y-enzyme reaction is enhanced by O2. Isoniazid-resistant mutants do not exhibit Y-enzyme, peroxidase or catalase activities, and do not take up isoniazid. The Y-enzyme reaction is therefore implicated in the uptake of the drug. 相似文献
11.
F J Fehrenbach 《Biochemical and biophysical research communications》1972,48(4):828-832
Treatment of red blood cells with NAD-glycohydrolase, streptolysin-O, EC 3.2.2.5, resulted in a complete loss of intracellular NAD. During the breakdown of NAD no appreciable hemolysis was observed. Cell lysis was detected first after complete destruction of NAD. It is suggested that enzyme molecules penetrated through the cellular membrane and that the enzymatic breakdown of cellular NAD is the key process in NAD-glycohydrolase induced hemolysis. 相似文献
12.
Analogues of oxidized nicotinamide adenine dinucleotide (NAD+) in which a 2,3-dihydroxycyclopentane ring replaces the beta-D-ribonucleotide ring of the nicotinamide riboside moiety of NAD+ have recently been synthesized [Slama, J. T., & Simmons, A. M. (1988) Biochemistry 27, 183]. Carbocyclic NAD+ analogues have been shown to inhibit NAD glycohydrolases and ADP-ribosyl transferases such as cholera toxin A subunit. In this study, the diastereomeric mixture of dinucleotides was separated, and the inhibitory capacity of each of the purified diastereomers was defined. The NAD+ analogue in which the D-dihydroxycyclopentane is substituted for the D-ribose is designated carba-NAD and was demonstrated to be a poor inhibitor of the Bungarus fasciatus venom NAD glycohydrolase. The diastereomeric dinucleotide pseudo-carbocyclic-NAD (psi-carba-NAD), containing L-dihydroxycyclopentane in place of the D-ribose of NAD+, was shown, however, to be a potent competitive inhibitor of the venom NAD glycohydrolase with an inhibitor dissociation constant (Ki) of 35 microM. This was surprising since psi-carba-NAD contains the carbocyclic analogue of the unnatural L-ribotide and was therefore expected to be a biologically inactive diastereomer. psi-Carba-NAD also competitively inhibited the insoluble brain NAD glycohydrolase from cow (Ki = 6.7 microM) and sheep (Ki = 31 microM) enzyme against which carba-NAD is ineffective. Sensitivity to psi-carba-NAD was found to parallel sensitivity to inhibition by isonicotinic acid hydrazide, another NADase inhibitor. psi-Carba-NAD is neither a substrate for nor an inhibitor of alcohol dehydrogenase, whereas carba-NAD is an efficient dehydrogenase substrate.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
13.
R J Mehta 《Canadian journal of microbiology》1977,23(4):402-406
An obligate methyltroph Methylomonas methylovora oxidized methylamine, formaldehyde, and formate. Enzymes oxidizing these substrates were detected in a cell-free system. Phenazine methosulfate-linked methylamine dehydrogenase was purified 21-fold. The enzyme had optimum activity at pH 7.5 and was stable at 60 degrees C for 5 min. The enzyme activity was inhibited by parachloromercuric benzoate, isonicotinic acid hydrazide, mercuric chloride, and sodium borate. 相似文献
14.
Autotrophically grown cells of Chloroflexus aurantiacus B-3 were shown to possess activity of ATP-dependent malate lyase (acetylating CoA). ATP: malate lyase is supposed to be the specific enzyme of the cycle of the autotrophic CO2 fixation, in which pyruvate synthase, pyruvate phosphate dikinase, phosphoenolpyruvate (PEP) carboxylase and malate dehydrogenase are involved as well. The main product of the CO2 fixation cycle is glyoxylate, which could further be converted into 3-phosphoglyceric acid (3-PGA) in the reactions of either glycerate or serine pathway. The enzymes of both pathways were detected in C. auratiacus B-3. The results of the in vivo studies of glyxoylate and glycine metabolism, as well as the inhibitor analysis using fluoroacetate (FAc), isonicotinic acid hydrazide (INH), and 4-aminopterin (4-AP) confirm the operation of the proposed pathway in Chloroflexus.Abbreviations 3-PGA
3-phosphoglyceric acid
- 4-AP
4-aminopterin
- FAc
fluoroacetate
- INH
isonicotinic acid hydrazide
- MV
methyl viologen
- PEP
phosphoenolpyruvate
- THF
tetrahydrofolate
- TPP
thiamine pyrophosphate 相似文献
15.
Purification and properties of an asparagine aminotransferase from Pisum sativum leaves 总被引:6,自引:0,他引:6
The enzyme responsible for the transamination of L-asparagine in pea leaves has been partially purified. It appears to be the same protein as the serine-glyoxylate aminotransferase. It is able to use serine or asparagine as amino donors and pyruvate or glyoxylate as amino acceptors. The reaction is reversible but the equilibrium is toward glycine or alanine production. The favored substrates are serine and glyoxylate: serine shows competitive inhibition toward asparagine, as does pyruvate toward glyoxylate. Substrate interaction and product inhibition patterns are consistent with a ping-pong mechanism. The enzyme has a pH optimum at 8.1. Gel filtration indicates a Mr of 105,000. Inhibition was caused by aminoxyacetate and hydroxylamine, but the enzyme was unaffected by isonicotinic acid hydrazide. The apoenzyme was resolved and was inactive: addition of pyridoxal 5'-phosphate restored 85% of the original activity. 相似文献
16.
NAD glycohydrolases are enzymes that catalyze the hydrolisis of NAD to produce ADP-ribose and nicotinamide. Regulation of these enzymes has not been fully elucidated. We have identified an NAD-glycohydrolase activity associated with the outer surface of the plasma membrane in human lung epithelial cell line A549. This activity is negatively regulated by its substrate -NAD but not by -NAD. Partial restoration of NADase activity after incubation of the cells with arginine or histidine, known ADP-ribose acceptors, suggests that inhibition be regulated by ADP-ribosylation. A549 do not undergo to apoptosis upon NAD treatment indicating that this effect be likely mediated by a cellular component(s) lacking in epithelial cells. 相似文献
17.
Nicotinamide adenine dinucleotide (NAD) content was found to decrease in mouse embryos during cleavage and to increase again at the blastocyst stage. The first enzyme involved in biosynthesis of NAD from nicotinamide is nicotinamide mononucleotide (NMN)-pyrophosphorylase. No such enzymatic activity was found in the embryos, but NAD-glycohydrolase activity was relatively high 24–48 hours after conception. Enzyme activity decreased in the blastocyst. The results are relevant to understanding the regulation of metabolism in preimplantation embryos. 相似文献
18.
The rate of incorporation of nicotinamide-[adenosine-U-14C]adenine dinucleotide [( Ado-U-14C]NAD) into histones and the poly(ADPR) polymerase activity of chromatin suggest that the NAD-dependent ADP-ribosylation of histones depends on the rate of NAD hydrolysis by glycohydrolase in chicken liver nuclei. With a rise in the NAD-glycohydrolase activity after treatment of nuclei with Triton X-100 the synthesis of poly(ADP-ribose) via the poly(ADPR)polymerase reaction is augmented, as a result of which the rate of [Ado-U-14C]NAD incorporation into total histones is increased. On the contrary, the decrease of NAD-glycohydrolase hydrolysis after treatment of nuclei with SDS lowers the poly(ADPR)polymerase activity and [Ado-U-14C]NAD incorporation into histones. Under these conditions, i. e. different rates of glycohydrolase hydrolysis of NAD in the nuclei, some redistribution of [Ado U-14C]NAD incorporation into individual histones occurs. 相似文献
19.
Radical scavenging activities of niacin-related compounds 总被引:4,自引:0,他引:4
Ogata S Takeuchi M Teradaira S Yamamoto N Iwata K Okumura K Taguchi H 《Bioscience, biotechnology, and biochemistry》2002,66(3):641-645
We investigated whether niacin-related compounds had radical-scavenging activity by electron spin resonance methods. Many compounds, but not trigonelline, had radical-scavenging activity against hydroxyl radicals. However, for the nitric oxide radical and 1,1-diphenyl-2-picrylhydrazyl radical, only nicotinic acid hydrazide and isonicotinic acid hydrazide had scavenging activities. These results suggest that the moiety of hydrazide might have an important role in scavenging abilities of various radicals. 相似文献
20.
Zikun Huang Qing Luo Yang Guo Jie Chen Guoliang Xiong Yiping Peng Jianqing Ye Junming Li 《PloS one》2015,10(6)
The tuberculous granuloma is an elaborately organized structure and one of the main histological hallmarks of tuberculosis. Macrophages, which are important immunologic effector and antigen-presenting cells, are the main cell type found in the tuberculous granuloma and have high plasticity. Macrophage polarization during bacterial infection has been elucidated in numerous recent studies; however, macrophage polarization during tuberculous granuloma formation and development has rarely been reported. It remains to be clarified whether differences in the activation status of macrophages affect granuloma formation. In this study, the variation in macrophage polarization during the formation and development of tuberculous granulomas was investigated in both sections of lung tissues from tuberculosis patients and an in vitro tuberculous granuloma model. The roles of macrophage polarization in this process were also investigated. Mycobacterium tuberculosis (M. tuberculosis) infection was found to induce monocyte-derived macrophage polarization. In the in vitro tuberculous granuloma model, macrophage transformation from M1 to M2 was observed over time following M. tuberculosis infection. M2 macrophages were found to predominate in both necrotic and non-necrotic granulomas from tuberculosis patients, while both M1 and M2 polarized macrophages were found in the non-granulomatous lung tissues. Furthermore, it was found that M1 macrophages promote granuloma formation and macrophage bactericidal activity in vitro, while M2 macrophages inhibit these effects. The findings of this study provide insights into the mechanism by which M. tuberculosis circumvents the host immune system as well as a theoretical foundation for the development of novel tuberculosis therapies based on reprogramming macrophage polarization. 相似文献