Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

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Query: EC:2.7.7.8 (polynucleotide phosphorylase)
723 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The inherited deficiency of purine-nucleoside phosphorylase (PNPase; purine-nucleoside:orthophosphate ribosyltransferase, EC 2.4.2.1) in humans is associated with a severe deficiency of the T lymphocytes of the immune system. Because of the unsatisfactory nature of previously described model systems, we have selected, cloned, and characterized a mutant mouse T cell lymphoma (S49) completely deficient in PNPase. Of the four substrates of PNPase, only deoxyguanosine at low concentrations is toxic to the PNPase-deficient (NSU-1) cells. In order to delineate the biochemical processes necessary for the sensitivity of the NSU-1 cells to deoxyguanosine, we have isolated a series of secondary mutants resistant to deoxyguanosine from the PNPase-deficient line. One of these mutants is defective in its ability to transport deoxyguanosine into the cell. A second type of mutant cannot phosphorylate the deoxyguanosine and is totally deficient in deoxycytidine kinase activity. A third type of mutant (NSU-1-dGuo-L) can both transport and phosphorylate deoxyguanosine and accumulates dGTP. However, unlike its parent, NSU-1-dGuo-L does not become depleted of dCTP and TTP when exposed to exogenous deoxyguanosine. This observation is accounted for by the fact that the reduction of CDP to dCDP by the ribonucleotide reductase (ribonucleoside-diphosphate reductase, 2'-deoxyribonucleoside-diphosphate:oxidized-thioredoxin 2'-oxidoreductase, EC 1.17.4.1) of NSU-1-dGuo-L cells is not normally sensitive to feedback inhibition by dGTP.Thus, in order to exert its toxicity deoxyguanosine must be transported into the cell, be phosphorylated by deoxycytidine kinase, and be accumulated as dGTP. By inhibiting ribonucleotide reductase, dGTP depletes the cell of dCTP and to some extent TTP, thus preventing the synthesis of DNA, a process necessary for any proliferation-dependent function of T cells.
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PMID:Isolation and characterization of purine-nucleoside phosphorylase-deficient T-lymphoma cells and secondary mutants with altered ribonucleotide reductase: genetic model for immunodeficiency disease. 10 75

The circulating lymphocytes of 16 normal subjects and of 18 patients with paraproteinemia have been characterized by measuring the levels of ADA, AMPA, PNPase, CDA. The results obtained reveal a highly significant increase in PNPase of subjects affected by paraproteinemia as compared to that of normal subjects (p less than 0.005). The fact that the increased enzymatic levels are found mainly in patients not affected by Bence-Jones proteinuria seems to indicate that the evolution of the paraproteinemia disease is in some way related to the intralymphocyte levels of PNPase.
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PMID:[Behavior of some enzymes of nucleoside catabolism in circulating lymphocytes during paraproteinemia]. 11 99

The reaction of 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole [NBD-Cl] with purified eel electrophax Na+ and K+ stimulated adenosine triphosphatase [(Na-K)ATPase] has been monitored by changes in the (Na-K)ATPase activity, the K+ stimulated p-nitrophenyl phosphatase [PNPase] activity, and the protein ultraviolet absorption spectrum. The NBD-Cl reacts with two tyrosine residues per mol of enzyme (approximately 6-7 nmol/mg of protein), as judged by changes in protein absorption spectra and incorporation of [14C]NBD-Cl. The modified tyrosine groups are located on the Mr = 95 000 polypeptide chain and react at different rates. Only one tyrosine modification is necessary for complete inhibition of (Na-K)ATPase activity, although both must be modified for complete inhibition of PNPase activity. Reversal of these modifications by 2-mercaptoethanol restores 65% of both activities. Na+ increases the rate of tyrosine modification, K+ decreases the rate, and ATP affords the more reactive tyrosine group complete protection. NBD-Cl modification of approximately 6-7 nmol of tyrosine groups/mg of protein results in a large decrease in ATP affinity as judged by equilibrium binding. These results are compared with similar results obtained from NBD-Cl modification of the coupling factors of oxidative phosphorylation and photophosphorylation. A model is presented suggesting an asymmetric arrangement of two 95 000 polypeptide chains with a single tyrosine residue at the ATP site.
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PMID:Reaction of (Na-K)ATPase with 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole: evidence for an essential tyrosine at the active site. 14 73

The E. coli polynucleotide phosphorylase-catalysed reaction of the deoxynucleoside 5'-diphosphates of 5-methyldeoxycytidine, N4-hydroxydeoxycytidine, deoxyuridine and 5-mercurideoxyuridine with the primers d(pT-T-A-G) and d(pT-T-T-T-T-T) have been studied under conditions where the primer is extended, predominantly, by one or two nucleotide residues. In experiments with 5-mercurideoxyuridine 5'-diphosphate, no 5-mercurideoxy-uridine-containing oligonucleotides were produced. The other three nucleotide analogs were found to be good substrates for E. coli PNPase and the conditions established for synthesis with these analogs will allow the construction of a number of biologically useful types of oligodeoxyribonucleotide.
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PMID:Enzymatic synthesis of oligodeoxyribonucleotides of defined sequence. Polynucleotide phosphorylase catalysed synthesis using pyrimidine analog-containing deoxyribonucleoside 5'-diphosphates. 35 64

Replication of RNA bacteriophages in the presence of rifamycin was studied in different Escherichia coli strains that vary in RNase content but are not isogenic: AB259 RNase+, Q13 RNase I- PNPase-, AB105 RNase I- RNase III-. It was found that rifamycin did not affect characteristics of phage replication such as the general pattern of viral RNA synthesis and intracellular development of the phage. These characteristics are strain specific and independent of the cell growth rate, which defines only phage release. The inhibition of cell division by rifamycin interfered with the release of the phage and thus produced an apparent effect of rifamycin on phage replication.
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PMID:Replication of RNA bacteriophages in the presence of rifamycin. 36 77

RNAase which usually contaminates commercial pancreatic DNAase preparations can be removed by affinity chromatography on agarose-coupled anti-RNAase antibodies. RNA treated with purified DNAase can be re-isolated intact, as determined by polyacrylamide gel electrophoresis under denaturing conditions. This method might be applicable to purification of other preparations which are used in RNA research, such as PNPase (polynucleotide phosphorylase) and specific antibodies for polysome immunoprecipitation. The non-specific binding of DNAase in our system is less than 5% and the loss of specific activity of DNAase I is less than 1%.
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PMID:A simple method for elimination of RNAase contamination from DNAase preparations. 55 95

On incubation of cells of E. coli B and MRE 600 (logariphmic phase of growth), treated with toluene in presence of a mixture 14C-nucleoside-5'-diphosphates, Mg2+ or Mn2+ and tris HCl buffer pH 8.0, intracellular synthesis of heteropolyribonucleotide was observed. The synthesis was catalyzed by polynucleotide phosphorylase (PNPase, E. C. 2.7.7.8). An increase in GDP concentration in the medium distinctly decreased the incorporation of other NDP into the polymer (poly-AGUC). If the ratio of ADP, UDP, CDP, GDP in the medium was 1:1:1:0.2, the composition of nitrogenous bases in the heteropolymer produced reflected completely the NDP concentrations in the incubation mixture. Addition of different amino acids (1-lysine, 1-histidine, glycine, 1-phenylalanine) and their mixtures stimulated poly-AGUC synthesis markedly and caused an appreciable alteration in the nucleotide composition of the poly-AGUC synthesized. This phenomenon resembled the effect of amino acids on the activity of partially purified PNPase and on RNA synthesis, catalized by the enzyme in vitro. These data suggest that in bacterial cell, i. e. in vivo, PNPase synthesizes specific RNA polyribonucleotide sequences, participating in protein synthesis or in its regulation.
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PMID:[Nucleotide composition of RNA, synthesized by polynucleotide phosphorylase, in toluene-treated cells of Escherichia coli]. 76 93

According to Cleland's theoretical predictions, inosine phosphorolysis catalyzed by chicken and pigeon's liver PNPase (Purine nucleoside:ortophosphate ribosyltransferase. E.C. 2.4.2.1.) appears to be a rapid equilibrium random bi-bi with "dead end" enzyme-phosphate-hypoxantine complex. This mechanism implies the existence of two essential active centers in the enzymatic molecule to which inosine and phosphate attach themselves independently. The observed lack of analogy in the PNPase mechanism of mechanism of different species seems to suggest the existence of structural differences between them.
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PMID:[Purine nucleoside phosphorylase. Catalytic reaction mechanism. II. Product-reaction-inhibition (author's transl)]. 81 90

Purified preparations of pigeon liver PNPase (E.C. 2.4.2.1) have been obtained by acid preparation of liver homogenates at pH = 5,followed by a fractionation with ammonium sulphate (25-50% saturation) and by a chromatographic adsorption on DEAE-cellulose. The preparation obtained shows a PNPase specific activity 325 times greater than that of the original homogenates. Kinetic studies carried out with homogenates and purified preparations of pigeon liver PNPase seem to suggest that inosine and deoxynosine react on the same catalytic site of the enzyme molecule.
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PMID:[Purification of purine nucleoside phosphorylase activity on deoxyinosine (author's transl)]. 82 97

In the phosphorolytic degradation catalyzed by chicken liver PNPase (E.C. 2.4.2.1) inosine appears to behave as a better substrate than xanthosine. Hypoxanthine, xanthine, guanine and purine (1 X 10(-1)M) appear to be inhibitors of the pigeon liver PNPase, whereas allopurinol, ATP, ITP, CTP and UTP (1. X 10(-3) M) do not inhibit the enzyme. Both PNPase activities exhibit the same optimum temperature (37-40 degrees C). Chicken liver PNPase optimum pH is in the range 6.5-7, whereas that of pigeon liver is in the range 7-7.5. Lineweaver-Burk plots for the inosine phosphorolysis catalyzed by chicken liver PNPase yielded straight lines if substrate concentrations were lower than 1 X 10(-4) M but concave downward curves at higher concentrations. This activation increases when the homogenates are stored at 4 degrees C and pH = 7 during 24 h or more; pigeon liver PNPase does not show this activation phenomenon.
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PMID:[Purine metabolites in the activity of purine nucleoside phosphorylase (author's transl)]. 82 98


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