Investigations on the role of Golgi-mediated, ligand-receptor processing in the activation of granulocytes by chemoattractants: differential effects of monensin.
Human granulocytes were exposed to different concentrations of the ionophore monensin for 20 min at 37 degrees C. Subsequent exposure to 50 nM of the chemoattractant fMet-Leu-[3H]Phe for up to 30 min at 37 degrees C resulted in a receptor-mediated uptake that was inhibited 80% at a monensin concentration of 30 microM. 50% inhibition was observed at 1-10 microM monensin with no significant change in fMet-Leu-Phe dose dependency. Subcellular fractionation of cells treated with monensin, indicated that the low density UDP-galactosyltransferase activity associated with internalized receptor-fMet-Leu-Phe complexes in untreated cells was absent. The high density galactosyltransferase activity cosedimenting with specific granule markers, however, was unaffected. Monensin also inhibited chemotaxis toward fMet-Leu-Phe as measured by migration of granulocytes through millipore filters and fMet-Leu-Phe induction of polarized morphology. Incubation of cell suspensions with up to 30 microM monensin, both before and during measurement of fMet-Leu-Phe stimulated superoxide production, did not affect the magnitude, kinetics, or transiency of the radical generation. Monensin did, however, shift the dose dependency of superoxide production of fMet-Leu-Phe to higher concentrations. These differential effects of monensin suggest that endocytosis of complexes of the chemoattractant and receptor is not involved in the activation or termination of the fMet-Leu-Phe stimulated superoxide production. They also are consistent with a role for receptor modulation and processing in the chemotactic response.
D002634:Chemotaxis, Leukocyte / Q000187:drug effects*; D006056:Golgi Apparatus / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure; D006098:Granulocytes / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure; D006801:Humans; D066298:In Vitro Techniques; D007700:Kinetics; D008854:Microscopy, Electron; D008985:Monensin / Q000494:pharmacology*; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000097:blood / Q000494:pharmacology*; D013481:Superoxides / Q000097:blood
D013481:Superoxides; D009240:N-Formylmethionine Leucyl-Phenylalanine; D008985:Monensin