ArticlePurinergic signalling2005
A capillary electrophoresis method for the characterization of ecto-nucleoside triphosphate diphosphohydrolases (NTPDases) and the analysis of inhibitors by in-capillary enzymatic microreaction.
Article in Purinergic signalling, 2005. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 27 papers.
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Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
27 citing papers in PubMed, 72 citations in OpenAlex.
- Heparins are potent inhibitors of ectonucleotide pyrophosphatase/phospho-diesterase-1 (NPP1) - a promising target for the immunotherapy of cancer.Frontiers in immunology · 2023Article
- Discovery of potent nucleotide pyrophosphatase/phosphodiesterase3 (NPP3) inhibitors with ancillary carbonic anhydrase inhibition for cancer (immuno)therapy.RSC medicinal chemistry · 2021Article
- Sulfated Polysaccharides from Macroalgae Are Potent Dual Inhibitors of Human ATP-Hydrolyzing Ectonucleotidases NPP1 and CD39.Marine drugs · 2021Article
- Development of Anthraquinone Derivatives as Ectonucleoside Triphosphate Diphosphohydrolase (NTPDase) Inhibitors With Selectivity for NTPDase2 and NTPDase3.Frontiers in pharmacology · 2020Article
- Salivary extracellular vesicles can modulate purinergic signalling in oral tissues by combined ectonucleoside triphosphate diphosphohydrolases and ecto-5'-nucleotidase activities.Molecular and cellular biochemistry · 2020Article
- Effects of the ecto-ATPase apyrase on microglial ramification and surveillance reflect cell depolarization, not ATP depletion.Proceedings of the National Academy of Sciences of the United States of America · 2018Article
- Review
- Rat submandibular glands secrete nanovesicles with NTPDase and 5'-nucleotidase activities.Purinergic signalling · 2015Article
- 8-BuS-ATP derivatives as specific NTPDase1 inhibitors.British journal of pharmacology · 2013Article
- Inhibition of ecto-ATPase activity by curcumin in hepatocellular carcinoma HepG2 cells.The journal of physiological sciences : JPS · 2012Article
- ATP hydrolysis pathways and their contributions to pial arteriolar dilation in rats.American journal of physiology. Heart and circulatory physiology · 2011Article
- Epitope mapping in cell surface proteins by site-directed masking: defining the structural elements of NTPDase3 inhibition by a monoclonal antibody.Protein engineering, design & selection : PEDS · 2010Article
- Synthesis of alkyl- and aryl-amino-substituted anthraquinone derivatives by microwave-assisted copper(0)-catalyzed Ullmann coupling reactions.Nature protocols · 2010Article
- Ectonucleotidases in the kidney.Purinergic signalling · 2009Article
- Role of ATP and related purines in inhibitory neurotransmission to the pig urinary bladder neck.British journal of pharmacology · 2009Article
- Structure-activity relationships of anthraquinone derivatives derived from bromaminic acid as inhibitors of ectonucleoside triphosphate diphosphohydrolases (E-NTPDases).Purinergic signalling · 2009Article
- Characterization of a monoclonal antibody as the first specific inhibitor of human NTP diphosphohydrolase-3 : partial characterization of the inhibitory epitope and potential applications.The FEBS journal · 2009Article
- Activity-dependent release of adenosine: a critical re-evaluation of mechanism.Current neuropharmacology · 2008Article
- Selective nucleoside triphosphate diphosphohydrolase-2 (NTPDase2) inhibitors: nucleotide mimetics derived from uridine-5'-carboxamide.Journal of medicinal chemistry · 2008Article
- Ectonucleotidases in Müller glial cells of the rodent retina: Involvement in inhibition of osmotic cell swelling.Purinergic signalling · 2007Article
Corrections and comments
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Authors and funding
5 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
A capillary electrophoresis (CE) method for the characterization of recombinant NTPDases 1, 2, and 3, and for assaying NTPDase inhibitors has been developed performing the enzymatic reaction within the capillary. After hydrodynamic injection of plugs of substrate solution with or without inhibitor in reaction buffer, followed by a suspension of an enzyme-containing membrane preparation, and subsequent injection of another plug of substrate solution with or without inhibitor, the reaction took place close to the capillary inlet. After 5 min, the electrophoretic separation of the reaction products was initiated by applying a constant current of -60 muA. The method employing a polyacrylamide-coated capillary and reverse polarity mode provided baseline resolution of substrates and products within a short separation time of less than 7 min. A 50 mM phosphate buffer (pH 6.5) was used for the separations and the products were detected by their UV absorbance at 210 nm. The Michaelis-Menten constants (K (m)) for the recombinant rat NTPDases 1, 2, and 3 obtained with this method were consistent with previously reported data. The inhibition studies revealed pronounced differences in the potency of reactive blue 2, pyridoxalphosphate-6-azophenyl-2',4'-disulfonic acid (PPADS), suramin, and N (6)-diethyl-beta,gamma-dibromomethylene-ATP (ARL67156) towards the NTPDase isoforms. Notably, ARL67156 does not inhibit all NTPDases, having only a minor inhibitory effect on NTPDase2. Dipyridamole is not an inhibitor of the NTPDase isoforms investigated. The new method is fast and accurate, it requires only tiny amounts of material (nanoliter scale), no sample pretreatment and can be fully automated; thus it is clearly superior to the current standard methods.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.