apyrase and Leukemia-P388

Tyrosine as well as serine/threonine protein kinase inhibitors have potentially two sites of interaction with their targets: the protein-substrate binding site and the ATP binding site. The latter could be modelized by measuring the capacity of protein kinase inhibitors to inhibit ATPase activities. In order to do so, we assess a novel, highly sensitive HPLC method based on hydrophilic separation of [gamma-32P]ATP and [32P]Pi. The novel assay is presented. Furthermore, the potency of 13 protein kinase inhibitors was tested on two types of ATPase, namely: apyrase and partially purified liver mitochondria F1-ATPase. The method described for the assay of ATPase can be used with almost any type of enzyme catalyzing this activity. Only cibacron blue and suramin show interesting capacities in inhibiting these ATPase activities pointing out that several widely used protein kinase inhibitors are at least somewhat specific in that they do not inhibit these two ATPases.

Topics: Adenosine Triphosphatases; Adenosine Triphosphate; Animals; Apyrase; Cell Membrane; Chromatography, High Pressure Liquid; Dose-Response Relationship, Drug; Kinetics; Leukemia P388; Mitochondria, Liver; Phosphates; Protein Kinase Inhibitors; Protein-Tyrosine Kinases; Proton-Translocating ATPases; Rats; Suramin; Triazines