Continuous fluorimetric assay of nucleotide pyrophosphatase. Kinetics, inhibitors, and extension to dinucleoside oligophosphatases
作者： Jacek Wierzchowski , Halina Sierakowska , David Shugar
DOI: 10.1016/0167-4838(85)90045-7
关键词: Enzyme kinetics 、 Substrate (chemistry) 、 Biochemistry 、 Kinetics 、 Hydrolysis 、 Fluorescence 、 Enzyme assay 、 Nucleotide pyrophosphatase 、 Chromatography 、 Enzyme 、 Chemistry
摘要: Abstract A simple and convenient procedure is described for the continuous fluorimetric assay of nucleotide pyrophosphatase (dinucleotide nucleotidohydrolase, EC 3.6.1.9) activity. It based on multifold increase in fluorescence emission intensity accompanying hydrolysis a new fluorogenic substrate, ϵAp 2 ϵA (where ϵ = 1, N 6 -etheno), or commercially available ϵNAD + FAD, using potato pyrophosphatase. The applicable to enzyme activity tissue extracts, as well kinetic studies screening potential inhibitors. K m V max / substrates, i values various compounds, were evaluated. most effective inhibitor was alternative substrate 8-bromo-NAD ( 7 μ M). method also useful snake venom phosphodiesterase I-5′-exo-nuclease (EC 3.1.4.1). preparation described, spectral properties substrates listed. Both 3 4 shown be I, may used specific dinucleoside oligophosphatases.
elsevier.com
sci-hub.se 参考文章(21)
W.E. Razzell, H.G. Khorana, Studies on polynucleotides. III. Enzymic degradation; substrate specificity and properties of snake venom phosphodiesterase. Journal of Biological Chemistry. ,vol. 234, pp. 2105- 2113 ,(1959) , 10.1016/S0021-9258(18)69874-6
C. V. BALAKRISHNAN, C. S. VAIDYANATHAN, N. Appaji RAO, Studies on nucleotidases in plants. Isolation and properties of the monomeric form of the crystalline and homogeneous mung bean nucleotide pyrophosphatase. FEBS Journal. ,vol. 78, pp. 95- 102 ,(1977) , 10.1111/J.1432-1033.1977.TB11717.X
J C Cameselle, M J Costas, M A Günther Sillero, A Sillero, Two low Km hydrolytic activities on dinucleoside 5‘,5“‘-P1,P4-tetraphosphates in rat liver. Characterization as the specific dinucleoside tetraphosphatase and a phosphodiesterase I-like enzyme. Journal of Biological Chemistry. ,vol. 259, pp. 2879- 2885 ,(1984) , 10.1016/S0021-9258(17)43230-3
A Ogilvie, W Antl, Diadenosine tetraphosphatase from human leukemia cells. Purification to homogeneity and partial characterization. Journal of Biological Chemistry. ,vol. 258, pp. 4105- 4109 ,(1983) , 10.1016/S0021-9258(18)32590-0
Rita VILLALBA, Alfredo MORENO, Miguel QUINTANILLA, Carmen D. LOBATON, Antonio SILLERO, Dinucleosidetriphosphatase from Rat Liver FEBS Journal. ,vol. 76, pp. 331- 337 ,(1977) , 10.1111/J.1432-1033.1977.TB11600.X
A Guranowski, H Jakubowski, E Holler, Catabolism of diadenosine 5',5"'-P1,P4-tetraphosphate in procaryotes. Purification and properties of diadenosine 5',5"'-P1,P4-tetraphosphate (symmetrical) pyrophosphohydrolase from Escherichia coli K12. Journal of Biological Chemistry. ,vol. 258, pp. 14784- 14789 ,(1983) , 10.1016/S0021-9258(17)43729-X
Marcjanna BARTKIEWICZ, Halina SIERAKOWSKA, David SHUGAR, Nucleotide pyrophosphatase from potato tubers: purification and properties FEBS Journal. ,vol. 143, pp. 419- 426 ,(1984) , 10.1111/J.1432-1033.1984.TB08389.X
J C Cameselle, M J Costas, M A G Sillero, A Sillero, Bis-(5'-guanosyl) tetraphosphatase in rat tissues. Biochemical Journal. ,vol. 201, pp. 405- 410 ,(1982) , 10.1042/BJ2010405
Malgorzata Zan-Kowalczewska, Marcianna Bartkiewicz, Halina Sierakowska, David Shugar, Purification and resolution of potato tuber cyclic nucleotide phosphodiesterase from nucleotide pyrophosphatase Biochimica et Biophysica Acta (BBA) - Protein Structure and Molecular Enzymology. ,vol. 788, pp. 62- 73 ,(1984) , 10.1016/0167-4838(84)90298-X
Nelson J. Leonard, Glen L. Tolman, FLUORESCENT NUCLEOSIDES AND NUCLEOTIDES Annals of the New York Academy of Sciences. ,vol. 255, pp. 43- 58 ,(1975) , 10.1111/J.1749-6632.1975.TB29212.X