PMID: 8452525Mar 1, 1993Paper

Kinetic characterization of guanine-nucleotide-induced exocytosis from permeabilized rat mast cells

The Biochemical Journal
T Lillie, B D Gomperts

Abstract

We have measured the time course of secretion of hexosaminidase from rat mast cells permeabilized (in simple buffered NaCl solutions) in response to guanine nucleotides [GTP or guanosine 5'-[gamma-thio]triphosphate (GTP[S])] and Ca2+. In these experiments, ATP was excluded from the system (and the cells were pretreated with metabolic inhibitors). For cells permeabilized in the absence of Mg2+ but in the presence of Ca2+, secretion commences promptly in response to addition of GTP; when Mg2+ (2 mM) is provided, secretion commences after an extended delay, much higher concentrations of GTP are required, and the final extent of secretion is decreased. Ongoing secretion due to GTP and Ca2+ is abruptly terminated by addition of Mg2+ to cells initially stimulated in its absence. In contrast, although Mg2+ has no effect on the sensitivity to the non-hydrolysable analogue GTP[S], its absence does nevertheless cause delays in the onset of secretion triggered by the addition of GTP[S] to cells initially permeabilized in the presence of Ca2+ (micromolar range, again in the absence of ATP). However, exocytosis from cells triggered with Ca2+ after permeabilization in the presence of high concentrations of GTP[S] is instantaneous. The delays...Continue Reading

Citations

Mar 29, 2003·Physiological Reviews·Robert D Burgoyne, Alan Morgan
Mar 9, 1994·Annals of the New York Academy of Sciences·B D GompertsP E Tatham
Jul 1, 1996·The American Journal of Physiology·J VadakekalamS A Metz
Apr 6, 1995·Biochimica Et Biophysica Acta·M MeredithS Metz
Apr 1, 1994·Neuron·J R Monck, J M Fernandez

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