Determination of Significance:
NADK (EC 2.7.1.23) is widely found in animals, plants, microorganisms and cultured cells. It is the only enzyme that can catalyze the phosphorylation of NAD+ to NADP+ in organisms. It can catalyze the phosphorylation reaction of NAD(H) with ATP or inorganic polyphosphate [poly(P)] as a phosphoryl donor to generate NADP(H). Therefore, NAD kinase plays an important role in the synthesis of NADP(H) and the regulation of the balance between NAD(H) and NADP(H).
Measurement Principle:
NADK catalyzes the phosphorylation of NAD+ to produce NADP+, which can be reduced to NADPH by glucose-6-phosphate dehydrogenase. NADPH could reduce oxidized thiazole blue (MTT) through the dehydrogenation of PMS. The absorbance value of MTT at 570 nm can be reflect the activity of NADK.
Self Provided:
Table centrifuge, spectrophotometer/microplate reader, water bath, micro glass cuvette/96 well flat –bottom plate (UV plate), adjustable pipette, mortar/homogenizer, anhydrous ethanol, ice and distilled water.
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