Enzyme Activity Measurement for Nucleoside-Diphosphate Kinase

Creative Enzymes is a world-wide leading company specialized in the development, optimization, and supply of analytical services. Well recognized in the global market, Creative Enzymes creates innovative enzyme activity assays that are performed with the most advanced instruments. The best technical support and personalized customer service are in the top class of the industry. Herein, we are proud to offer the reliable enzyme activity assay for nucleoside-diphosphate kinase.

The family of nucleoside-diphosphate kinases (EC 2.7.4.6; ATP:nucleoside-diphosphate phosphotransferase) consists of a specific group of ubiquitous housekeeping phosphotransferases. They are implicated in maintaining the pool of intracellular nucleoside triphosphates (NTPs) required for biosynthesis by catalyzing the non-substrate-specific conversion of nucleosides diphospate (NDP) to NTP. Particularly, nucleoside-diphosphate kinase catalyzes the reversible transfer of a terminal γ-phosphoryl group (P) from NTP to NDP involving a high-energy phosphorylated and conserved histidine residue (His) as intermediate, by means of a ping-pong mechanism. Nucleoside-diphosphate kinases have been isolated from various organisms and cell types, including prokaryotes, lower and higher eukaryotes, mouse, rat, and human. They are ubiquitously distributed, particularly in the nucleus, in the cytoplasm associated to the microtubules, on the cell surface, in the mitochondria, and in the extracellular fluid. Eukaryotic nucleoside-diphosphate kinases are homohexamers, whereas bacterial nucleoside-diphosphate kinases are homotetramers.

Nucleoside-diphosphate kinases play a critical role in many aspects: (1) the enzymes can use ATP as a donor to synthesize the nonadenylic NTPs needed for nucleic acid synthesis and several important metabolic intermediates including UDP-glucose and a few CDP-lipid derivatives; (2) the enzymes catalyze transphosphorylation between GTP produced in the Krebs cycle and ADP; (3) the enzymes can provide GTP for protein synthesis, G-protein signaling, and tubulin polymerization. Additionally, nucleoside-diphosphate kinases have been found in the biological regulation of growth, development, apoptosis, and differentiation. Moreover, the enzymes can act as the suppression of metastasis in tumors, the activity of histidine-dependent protein kinase, and the 3’-5’ exonuclease activity. Furthermore, nucleoside-diphosphate kinases also promote tumor formation and play multiple roles in normal development and cellular proliferation. Therefore, nucleoside-diphosphate kinases are attracting an ever-increasing interest for their various significant functions.

Creative Enzymes is capable of performing the most precise enzymatic assay for nucleoside-diphosphate kinases. The activity of nucleoside-diphosphate kinases is determined spectrophotometrically by a coupled assay. ATP formation by the kinase reaction is coupled to the reduction of NADP⁺ (or ADP formation was coupled to NADH oxidation), and the progress of the reaction is followed by monitoring the change in absorbance at 340 nm. The quality of our results is assured in a professional and timely manner, which is backed by the most advanced spectrophotometric instruments, the leading techniques, and a superb expert team. Ultimately, Creative Enzymes is your optimal choice for any research involving nucleoside-diphosphate kinases.

Figure: The crystal structure of nucleoside-diphosphate kinase from Dictyostelium discoideum.
PDB: 1PAE