Features of the translocation of protein kinase C in neutrophils stimulated with the chemotactic peptide f-Met-Leu-Phe.

The effects of f-Met-Leu-Phe (fMLP) on neutrophils, i.e. elevation of the levels of cytoplasmic Ca2+ and intramembranous diacylglycerol, would be expected to be accompanied by translocation of protein kinase C (PKC) to the plasmalemma. However, fMLP-induced PKC translocation could hitherto be demonstrated only when cells were additionally treated with cytochalasin B. We show here that treatment of guinea pig neutrophils with fMLP alone does lead to a significant PKC translocation which can be inhibited by pertussis toxin. The translocation can be detected only if the incubation is terminated within 30 sec after addition of fMLP, the termination is rapid, e.g. by application of a freeze clamp-technique, and the concentration of Ca2+ chelators in the buffer used for lysing the cells is low.