GTP-induced fusion of isolated pancreatic microsomal vesicles is increased by acidification of the vesicle lumen.

TY - JOUR

T1 - GTP-induced fusion of isolated pancreatic microsomal vesicles is increased by acidification of the vesicle lumen.

AU - Hampe, Wolfgang

AU - Zimmermann, P

AU - Schulz, I

PY - 1990

Y1 - 1990

N2 - Using the 'fusogen' polyethyleneglycol (PEG), Dawson et al. have concluded that both guanosine triphosphate (GTP)-induced calcium efflux and the enhancement of IP3-promoted calcium release from rat liver microsomal vesicles could be attributed to a GTP-dependent vesicle fusion. We have studied GTP-induced fusion of microsomal vesicles from rat exocrine pancreas using light scatter and fluorescence dequenching methods. In the presence of PEG (3%), GTP (10 microM) induced a decrease in light scatter and an increase in fluorescence in the fluorescence dequenching assay (GTP-effect) indicating fusion of the vesicles. Guanosine 5'-O-(3-thiotriphosphate) (10 microM) had no effect on its own and inhibited the GTP-induced signals. Preincubation of the vesicles with adenosine triphosphate (ATP) (4 mM) increased the GTP-effect by 80%, whereas bafilomycin B1, a specific inhibitor of vacuolar type H(+)-ATPases, and the protonophore CCCP (10 microM) inhibited only the ATP-dependent part of the GTP-effect. Inhibitors of the vacuolar type H(+)-ATPase, which are also SH-alkylating reagents such as N-ethylmaleimid (100 microM) and the tyrosine-, cysteine- and lysine-reactive reagent 7-chloro-4-nitrobenz-2-exa-1,3-diazole (10 microM), abolished the GTP-effect in the absence or presence of ATP. We conclude that GTP induces fusion of pancreatic microsomes which is increased by an H+ gradient established by a vacuolar type H(+)-ATPase.

AB - Using the 'fusogen' polyethyleneglycol (PEG), Dawson et al. have concluded that both guanosine triphosphate (GTP)-induced calcium efflux and the enhancement of IP3-promoted calcium release from rat liver microsomal vesicles could be attributed to a GTP-dependent vesicle fusion. We have studied GTP-induced fusion of microsomal vesicles from rat exocrine pancreas using light scatter and fluorescence dequenching methods. In the presence of PEG (3%), GTP (10 microM) induced a decrease in light scatter and an increase in fluorescence in the fluorescence dequenching assay (GTP-effect) indicating fusion of the vesicles. Guanosine 5'-O-(3-thiotriphosphate) (10 microM) had no effect on its own and inhibited the GTP-induced signals. Preincubation of the vesicles with adenosine triphosphate (ATP) (4 mM) increased the GTP-effect by 80%, whereas bafilomycin B1, a specific inhibitor of vacuolar type H(+)-ATPases, and the protonophore CCCP (10 microM) inhibited only the ATP-dependent part of the GTP-effect. Inhibitors of the vacuolar type H(+)-ATPase, which are also SH-alkylating reagents such as N-ethylmaleimid (100 microM) and the tyrosine-, cysteine- and lysine-reactive reagent 7-chloro-4-nitrobenz-2-exa-1,3-diazole (10 microM), abolished the GTP-effect in the absence or presence of ATP. We conclude that GTP induces fusion of pancreatic microsomes which is increased by an H+ gradient established by a vacuolar type H(+)-ATPase.

M3 - SCORING: Zeitschriftenaufsatz

VL - 271

SP - 62

EP - 66

JO - FEBS LETT

JF - FEBS LETT

SN - 0014-5793

IS - 1-2

M1 - 1-2

ER -