Direct and indirect receptor‐independent G‐protein activation by cationic‐amphiphilic substances. Studies with mast cells, HL‐60 human leukemic cells and purified G‐proteins

Studies from several laboratories have revealed that structurally diverse substances including the wasp venom, mastoparan (MP), activate purified regulatory heterotrimeric guanine nucleotidc-binding proteins (G-proteins) in a receptor-independent manner, presumably by mimicking the effects of heptahelical receptors. Mast cells and differentiated HL-60 human leukemic cells are useful model systems for the analysis of receptor-independent G-protein activation. We compared the effects of 2-phenylhistamines which are cationic-amphiphilic, too, and of MP on G-protein activation in dibutyryl cAMP-differentiated HL-60 cells and in the rat basophilic leukemia cell line, RBL 2H3. In HL-60 cells, 2-phenylhistamines show stimulatory effects which resemble those of formyl peptide receptor agonists but which cannot be attributed to agonism at classical receptors. 2-phenylhistamines do not, however, activate RBL 2H3 cells and various other myeloid cell types, pointing to cell type-specificity of receptor-independent G-protein activation. In HL-60 cells, MP shows effects on G-protein activation which differ substantially from those of formyl peptides. In RBL 2H3 membranes, MP shows similar effects on G-prolein activation as in HL-60 membranes. We develop a model according to which receptor-independent G-protein activation can be subdivided into direct and indirect receptor-independent G-protein activation. In case of the former mechanism, substances like 2-phenylhislamines interact with G-protein α-subunits and in case of the latter mechanism, substances like MP interact with nucleoside diphosphate kinase which catalyzes the formation of GTP. This newly formed GTP is then transferred to, and cleaved by, G-protein a-subunits. NDPK is a novel target for the design of drugs which interfere with G-protein-mcdiated signal transduction at a post-receptor level and may modulate the function of various cell types including mast cells.