Inhibition by cromoglycate and some flavonoids of nucleoside diphosphate kinase and of exocytosis from permeabilized mast cells.
1. The anti-allergic compound, cromoglycate, is reported to possess affinity for, and to suppress the autophosphorylation of a 72kDa protein having the sequence of nucleoside diphosphate kinase (NDPK). 2. We have tested the ability of cromoglycate, and a panel of ten structurally related flavonoids of plant origin, to inhibit the NDPK reaction and the exocytotic process of permeabilized mast cells. The conditions of permeabilization (use of an isotonic medium based on sodium glutamate) were selected to ensure that NDPK activity would be an essential component in the induction of Ca(2+)-induced exocytosis in which ATP is required for generation of GTP. For comparison, we also measured the inhibition of exocytosis induced by GTP-gamma-S; this proceeds in the absence of ATP and bypasses the need for NDPK activity. 3. We found that cromoglycate does not discriminate between Ca2+ and GTP-gamma-S-induced exocytosis and is a poor inhibitor of NDPK activity. Concentrations in the millimolar range are required for inhibition of all these functions. By comparison, many of the flavonoids are effective at concentrations in the micromolar range. 4. While we were unable to discern any systematic relationships between their ability to inhibit the three functions, two compounds, quercetin and genistein, inhibit Ca(2+)-induced, but not GTP-gamma-S-induced exocytosis. Inhibition of the late stages of the stimulus-response pathway in mast cells by these compounds is therefore likely to be due to inhibition of NDPK and the consequent failure to generate GTP.
Martin
M W
MW
Department of Physiology, University College London.
O'Sullivan
A J
AJ
Gomperts
B D
BD
D000818:Animals; D004205:Cromolyn Sodium / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D005089:Exocytosis / Q000187:drug effects*; D005419:Flavonoids / Q000494:pharmacology*; D008297:Male; D008407:Mast Cells / Q000187:drug effects*; D009701:Nucleoside-Diphosphate Kinase / Q000187:drug effects*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013997:Time Factors
D005419:Flavonoids; D009701:Nucleoside-Diphosphate Kinase; D004205:Cromolyn Sodium