A high-performance liquid chromatographic method for the assay of Na+,K+-adenosine triphosphatase inhibition.
- 1 January 1985
- journal article
- research article
- Published by Pharmaceutical Society of Japan in Journal of Pharmacobio-Dynamics
- Vol. 8 (1) , 64-68
- https://doi.org/10.1248/bpb1978.8.64
Abstract
A new method for the assay of Na+,K+-ATPase inhibition was devised involving the determination of enzymatically produced ADP and unchanged ATP by high-performance liquid chromatography (HPLC). The substrate, ATP, was incubated with the enzyme preparation in the presence of an inhibitor. The incubation mixture was filtered through a membrane filter, and ADP and ATP in the filtrate were separated by (HPLC). The inhibitory effect of a cardiac steroid on the enzymic reaction was estimated by measuring the peak area ratio of ADP to ADP plus ATP on the chromatogram. The proposed assay method has proved to be satisfactory with respect to simplicity, sensitivity and reproducibility.This publication has 7 references indexed in Scilit:
- Heterogeneity of ouabain specific binding sites and (Na+ + K+)-ATPase inhibition in microsomes from rat heartBiochemical Pharmacology, 1984
- A volatile liquid chromatography system for nucleotidesAnalytical Biochemistry, 1981
- Inhibition by digoxin and SC4453 of (Na+ + K+)-ATPase prepared from human heart, guinea-pig heart and guinea-pig brainEuropean Journal of Pharmacology, 1979
- Studies on cardiotonic steroids from the skin of Japanese toad.CHEMICAL & PHARMACEUTICAL BULLETIN, 1976
- Membrane Adenosine Triphosphatase as a Participant in the Active Transport of Sodium and Potassium in the Human ErythrocyteJournal of Biological Chemistry, 1960
- Further investigations on a Mg++ + Na+-activated adenosintriphosphatase, possibly related to the active, linked transport of Na+ and K+ across the nerve membraneBiochimica et Biophysica Acta, 1960
- PROTEIN MEASUREMENT WITH THE FOLIN PHENOL REAGENTJournal of Biological Chemistry, 1951