[CITATION][C] Determination of phosphoenolpyruvate carboxykinase activity with deoxyguanosine 5′-diphosphate as nucleotide substrate
I Petrescu, O Bojan, M Saied, O Barzu, F Schmidt… - Analytical …, 1979 - Elsevier
I Petrescu, O Bojan, M Saied, O Barzu, F Schmidt, HF Kühnle
Analytical biochemistry, 1979•ElsevierNTP+ oxalacetate c NDP+ phosphoenolpyruvate+ COZ, where NTP and NDP stand for
nucleoside tri-and diphosphate, respectively. The mammalian enzyme has a rather high
degree of specificity for nucleotides, accepting as substrates only inosine, guanosine, and
xanthosine nucleotides (1, 2). The common features of these compounds are a keto group at
C, and an NH group at N, of the purine ring. The PEPCK activity is usually assayed by
measuring the phosphoenolpyruvate production in the forward reaction (3, 4) or the fixation …
nucleoside tri-and diphosphate, respectively. The mammalian enzyme has a rather high
degree of specificity for nucleotides, accepting as substrates only inosine, guanosine, and
xanthosine nucleotides (1, 2). The common features of these compounds are a keto group at
C, and an NH group at N, of the purine ring. The PEPCK activity is usually assayed by
measuring the phosphoenolpyruvate production in the forward reaction (3, 4) or the fixation …
NTP+ oxalacetate c NDP
+ phosphoenolpyruvate+ COZ, where NTP and NDP stand for nucleoside tri-and diphosphate, respectively. The mammalian enzyme has a rather high degree of specificity for nucleotides, accepting as substrates only inosine, guanosine, and xanthosine nucleotides (1, 2). The common features of these compounds are a keto group at C, and an NH group at N, of the purine ring.
The PEPCK activity is usually assayed by measuring the phosphoenolpyruvate production in the forward reaction (3, 4) or the fixation of carbon dioxide in the backward reaction (56). As the procedures consist of several steps including protein precipitation, chromatographic separation of reactants or radioisotope determination, our aim was to find a more rapid and specific method, suitable for a large number of measurements. A very convenient procedure is to couple the formation of oxalacetate in the backward reaction with NADH
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