Development of an enzyme cycling method by a purine nucleoside phosphorylase for assaying inorganic phosphate

2017 ◽  
Vol 9 (44) ◽  
pp. 6235-6239 ◽  
Author(s):  
Shigeru Ueda ◽  
Shin-ichi Sakasegawa
Keyword(s):  
Inorganic Phosphate ◽  
Purine Nucleoside Phosphorylase ◽  
Purine Nucleoside ◽  
Nucleoside Phosphorylase ◽  
Enzymatic Cycling

We have developed a novel enzymatic cycling method that uses purine nucleoside phosphorylase (PNP) (EC 2.4.2.1) fromBacillussp. to measure inorganic phosphate.

Purine Nucleoside Phosphorylase: Its Use in a Spectroscopic Assay for Inorganic Phosphate and for Removing Inorganic Phosphate with the Aid of Phosphodeoxyribomutase

1998 ◽  
Vol 265 (2) ◽  
pp. 299-307 ◽  
Author(s):  
Andrew E. Nixon ◽  
Jackie L. Hunter ◽  
Gordon Bonifacio ◽  
John F. Eccleston ◽  
Martin R. Webb
Keyword(s):  
Inorganic Phosphate ◽  
Purine Nucleoside Phosphorylase ◽  
Purine Nucleoside ◽  
Nucleoside Phosphorylase

Purine Nucleoside Phosphorylase as a Molecular Target to Develop Active Compounds Against Mycobacterium Tuberculosis

2010 ◽  
Vol 1 (1) ◽  
pp. 34
Author(s):  
Rodrigo G. Ducati ◽  
André A. Souto ◽  
Rafael A. Caceres ◽  
Walter F. De Azevedo ◽  
Luiz A. Basso ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Purine Nucleoside Phosphorylase ◽  
Molecular Target ◽  
Purine Nucleoside ◽  
Active Compounds ◽  
Nucleoside Phosphorylase

Substrate Specificity of a Purine Nucleoside Phosphorylase from Aeromonas hydrophila Toward 6-Substituted Purines and its Use as a Biocatalyst in the Synthesis of the Corresponding Ribonucleosides

2015 ◽  
Vol 19 (22) ◽  
pp. 2220-2225 ◽  
Author(s):  
Daniela Ubiali ◽  
Carlo Morelli ◽  
Marco Rabuffetti ◽  
Giulia Cattaneo ◽  
Immacolata Serra ◽  
...  
Keyword(s):  
Substrate Specificity ◽  
Aeromonas Hydrophila ◽  
Purine Nucleoside Phosphorylase ◽  
Purine Nucleoside ◽  
Nucleoside Phosphorylase

Purine nucleoside phosphorylase: Isolation and characterization

1990 ◽  
Vol 55 (12) ◽  
pp. 2987-2999 ◽  
Author(s):  
Katarina Šedivá ◽  
Ivan Votruba ◽  
Antonín Holý ◽  
Ivan Rosenberg
Keyword(s):  
Molecular Weight ◽  
Purine Nucleoside Phosphorylase ◽  
Purine Nucleoside ◽  
Leukemia Cells ◽  
Ph Optimum ◽  
Nucleoside Phosphorylase ◽  
Isolation And Characterization ◽  
Reaction Proceeds ◽  
The Matrix ◽  
Sepharose 4B

Purine nucleoside phosphorylase (PNP) from mouse leukemia cells L1210 was purified to homogeneity by a combination of ion exchange and affinity chromatography using AE-Sepharose 4B and 9-(p-succinylaminobenzyl)hypoxanthine as the matrix and the ligand, respectively. The native enzyme has a molecular weight of 104 000 and consists of three subunits of equal molecular weight of 34 000. The results of isoelectric focusing showed that the enzyme is considerably microheterogeneous over the pI-range 4.0-5.8 and most likely consists of eight isozymes. The temperature and pH-optimum of phosphorolysis, purine nucleoside synthesis and also of transribosylation is identical, namely 55 °C and pH 7.4. The transribosylation reaction proceeds in the presence of phosphate only. The following Km-values (μmol l-1) were determined for phosphorolysis: inosine 40, 2'-deoxyinosine 47, guanosine 27, 2'-deoxyguanosine 32. The Km-values (μmol l-1) of purine riboside and deoxyriboside synthesis are lower than the values for phosphorolysis (hypoxanthine 18 and 34, resp., guanine 8 and 11, resp.). An affinity lower by one order shows PNP for (-D-ribose-1-phosphate, (-D-2-deoxyribose-1-phosphate (Km = 200 μmol l-1 in both cases) and phosphate (Km = 805 μmol l-1). The substrate specificity of the enzyme was also studied: positions N(1), C(2) and C(8) are decisive for the binding of the substrate (purine nucleoside).

Purine Nucleoside Phosphorylase (PNP) Deficiency Leading to Accumulation of Lymphocytes in S-Phase

1986 ◽  
Vol 3 (4) ◽  
pp. 353-359 ◽  
Author(s):  
Ger T. Rijkers ◽  
Ben J. M. Zegers ◽  
Leo J. M. Spaapen ◽  
Derk H. Rutgers ◽  
John J. Roord ◽  
...  
Keyword(s):  
Purine Nucleoside Phosphorylase ◽  
S Phase ◽  
Purine Nucleoside ◽  
Nucleoside Phosphorylase
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