A cell-physiological description of GE, a GTP-binding protein that mediates exocytosis.
Introduction of GTP gamma S or other non-metabolic analogues of GTP into permeabilized myeloid granulocytes (mast cells, eosinophils, neutrophils) constitutes a sufficient stimulus to induce exocytosis. We concentrate on mast cells. Exocytosis from cells permeabilized in isotonic glutamate solution proceeds in the absence of ATP and at exceedingly low levels (< 10(-9) M) of Ca2+. Mg2+ strongly promotes GTP gamma S-induced exocytosis but this requirement can be spared and then obliterated by lifting Ca2+ through 10(-7) to 10(-6) M. GTP provides only a modest support to exocytosis but becomes almost equipotent with GTP gamma S when Mg2+ is excluded. Ca2+ alone is unable to induce exocytosis. We envisage that the terminal stage of exocytosis (membrane fusion) requires activation of GE, a putative GTPase so far undefined as a molecular entity. Ca2+, presumed to act through a Ca(2+)-binding protein (CE, also undefined) supports exocytosis by promoting the exchange of guanine nucleotides on GE. In the absence of Mg2+ the onset of exocytosis is characterized by delays that have concentration-dependent (binding) and independent components. The latter are sensitive to the identity of the stimulating nucleotide (GTP < GTP gamma S < Gpp [NH]p) and may reflect activation of GE. The activation by Ca2+ and Mg2+ and the delays preceding onset of GTP gamma S-triggered exocytosis are reminiscent of the action of glucagon and Mg2+ in the activation of adenylate cyclase in hepatocyte membranes. The cell-physiological description predicts GE to be an alpha beta gamma heterotrimeric GTP-binding protein with functional similarity to GS.
Lillie
T H
TH
Department of Physiology, University College London, UK.
Gomperts
B D
BD
D002118:Calcium / Q000502:physiology; D005089:Exocytosis / Q000502:physiology*; D020558:GTP Phosphohydrolases / Q000502:physiology; D019204:GTP-Binding Proteins / Q000502:physiology*; D006160:Guanosine Triphosphate / Q000502:physiology; D066298:In Vitro Techniques; D007700:Kinetics
D006160:Guanosine Triphosphate; D020558:GTP Phosphohydrolases; D019204:GTP-Binding Proteins; D002118:Calcium