Subcellular fractionation of human neutrophils on linear sucrose density gradients was utilized to test the hypothesis that priming regulates the subcellular and sub-plasma membrane distribution of neutrophil G-protein subunits, G(ialpha2) and G(ialpha3), N-formyl peptide receptor, Lyn kinase and phospholipase C-beta2. G(ialpha2), but not G(ialpha3), moved from a lighter to a higher density plasma membrane fraction. Unoccupied N-formyl peptide receptors were found throughout the plasma membrane fractions and this distribution did not change with priming. In imprinted cells G(ialpha2) and its effector, phospholipase C-beta2, were segregated in different membrane compartments; priming caused G(ialpha2) to move to the compartment in which phospholipase C-beta2 resided. Thus, an important component of the mechanism of priming may involve regulation of the location of G-protems and effector molecules in plasma membrane compartments where their abilities to couple may be enhanced. (C) 2003 Elsevier Science (USA). All rights reserved.