ArticlePloS one2011
Phosphatase-coupled universal kinase assay and kinetics for first-order-rate coupling reaction.
Article in PloS one, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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Who cites it
12 citing papers in PubMed, 26 citations in OpenAlex.
- Optimization of a malachite green-pyrophosphatase assay for the Zika virus RNA-dependent RNA polymerase and preliminary identification of natural product modulators.Frontiers in drug discovery · 2026Article
- Mass spectrometry-based phosphoproteomics in clinical applications.Trends in analytical chemistry : TRAC · 2023Article
- 5-Ethynyl-2'-deoxycytidine and 5-ethynyl-2'-deoxyuridine are differentially incorporated in cells infected with HSV-1, HCMV, and KSHV viruses.The Journal of biological chemistry · 2020Article
- A Ribose-Scavenging System Confers Colonization Fitness on the Human Gut Symbiont Bacteroides thetaiotaomicron in a Diet-Specific Manner.Cell host & microbe · 2020Article
- High level in vivo mucin-type glycosylation in Escherichia coli.Microbial cell factories · 2018Article
- A generic HTS assay for kinase screening: Validation for the isolation of an engineered malate kinase.PloS one · 2018Article
- Characterisation of the selective binding of antibiotics vancomycin and teicoplanin by the VanS receptor regulating type A vancomycin resistance in the enterococci.Biochimica et biophysica acta. General subjects · 2017Article
- Structural insights into mis-regulation of protein kinase A in human tumors.Proceedings of the National Academy of Sciences of the United States of America · 2015Article
- Expression of the functional recombinant human glycosyltransferase GalNAcT2 in Escherichia coli.Microbial cell factories · 2015Article
- Monitoring ATP hydrolysis and ATPase inhibitor screening using (1)H NMR.Chemical communications (Cambridge, England) · 2014Article
- Comparative Structural and Functional Analysis of Staphylococcus aureus Glucokinase with other Bacterial Glucokinases.Indian journal of pharmaceutical sciences · 2014Article
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Authors and funding
1 author at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Kinases use adenosine-5'-triphosphate (ATP) as the donor substrate and generate adenosine-5'-diphosphate (ADP) as a product. An ADP-based phosphatase-coupled kinase assay is described here. In this assay, CD39L2, a nucleotidase, is added into a kinase reaction to hydrolyze ADP to AMP and phosphate. The phosphate is subsequently detected using malachite green phosphate-detection reagents. As ADP hydrolysis by CD39L2 displays a first-order rate constant, relatively simple equations are derived to calculate the coupling rate and the lagging time of the coupling reaction, allowing one to obtain kinase kinetic parameters without the completion of the coupling reaction. ATP inhibition of CD39L2-catalyzed ADP hydrolysis is also determined for correction of the kinetic data. As examples, human glucokinase, P. chrysogenum APS kinase and human ERK1, kinases specific for sugar, nucleotide and protein respectively, are assayed. To assess the compatibility of the method for high-throughput assays, Z' factors >0.5 are also obtained for the three kinases.
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