Inhibition of exocytosis by intracellularly applied antibodies against a chromaffin granule-binding protein.
Exocytotic secretion requires the interaction and fusion of secretory vesicles with the plasma membrane. This process could be mediated by specific recognition molecules acting as intracellular, membrane-bound receptors and ligands. One possible component of such a recognition site on the plasma membrane is a protein of relative molecular mass (Mr) 51,000 (51K) that has been isolated from bovine adrenal chromaffin cells. This protein binds strongly to chromaffin granules, the secretory vesicles of these cells. To determine the function of this membrane-anchored chromaffin granule-binding protein in exocytosis, we tested the effect of intracellularly injected antibodies on secretion. Here we show, by two independent techniques in two different cell types, that antibodies against this protein inhibit exocytosis. In rat pheochromocytoma cell cultures, monospecific antibodies, applied by erythrocyte ghost fusion, impair the release of 3H-noradrenaline. The same antibodies, introduced into individual chromaffin cells through a patch pipette, block exocytosis, as revealed by the measurement of membrane capacitance. These results demonstrate the functional involvement in exocytosis of a plasma membrane protein with high affinity for secretory vesicles.
Schweizer
F E
FE
Friedrich Miescher Institut, Basel, Switzerland.
Schäfer
T
T
Tapparelli
C
C
Grob
M
M
Karli
U O
UO
Heumann
R
R
Thoenen
H
H
Bookman
R J
RJ
Burger
M M
MM
D000313:Adrenal Medulla / Q000502:physiology; D000818:Animals; D000906:Antibodies / Q000302:isolation & purification; D000936:Antigen-Antibody Complex; D002352:Carrier Proteins / Q000276:immunology / Q000302:isolation & purification / Q000502:physiology*; D002417:Cattle; D002460:Cell Line; D002462:Cell Membrane / Q000502:physiology; D002478:Cells, Cultured; D002837:Chromaffin Granules / Q000502:physiology*; D002838:Chromaffin System / Q000502:physiology*; D005089:Exocytosis; D008970:Molecular Weight; D011480:Protease Inhibitors / Q000494:pharmacology*
D000906:Antibodies; D000936:Antigen-Antibody Complex; D002352:Carrier Proteins; D011480:Protease Inhibitors