Detection of G-protein heterotrimers on large dense core and small synaptic vesicles of neuroendocrine and neuronal cells.
Heterotrimeric G proteins, initially believed to be exclusively present in the plasma membrane, have also been found to be associated with intracellular membrane compartments. There they are involved in various membrane trafficking processes including regulated secretion (reviewed in Bomsel, M., K. Mostov, Mol. Biol. Cell 3, 1317-1328 (1992)). Vesicles of two distinct types enter the regulated secretory pathway, i.e. large dense core vesicles and small synaptic vesicles, which differ in their membrane composition and content. Little is known about an association of heterotrimeric G proteins with regulated secretory vesicles, that would explain some aspects of the role heterotrimeric G proteins have during secretion. By immunofluorescence microscopy and immunoreplica analysis, we provide the first demonstration of the presence of complete sets of heterotrimeric G proteins, consisting of alpha-, beta-, and gamma-subunits, on large dense core vesicles from bovine adrenal medulla (chromaffin granules) and small synaptic vesicles from rodent and bovine brain. Each of the two types of secretory vesicles contains beta-subunits (at least beta 1 and beta 2), as well as gamma-subunits (at least gamma 2 or gamma 3). Interestingly, they differ in their composition of alpha-subunits. On small synaptic vesicles, we found two G(o) alpha-subunits (alpha o1 and alpha o2) and two Gi alpha-subunits (alpha i1 and alpha i2). In contrast, on chromaffin granules so far only one alpha o-subunit but no alpha i-subunits could be detected. Functional properties such as transmitter storage and/or exocytotic membrane fusion may be modulated by the various G-protein subunits associated with chromaffin granules and small synaptic vesicles.
Ahnert-Hilger
G
G
Abteilung Gastroenterologie, Universitätsklinikum Steglitz, Freie Universität, Berlin/Germany.
Schäfer
T
T
Spicher
K
K
Grund
C
C
Schultz
G
G
Wiedenmann
B
B
D000313:Adrenal Medulla / Q000737:chemistry* / Q000648:ultrastructure; D000818:Animals; D000918:Antibody Specificity; D002417:Cattle; D002478:Cells, Cultured; D019204:GTP-Binding Proteins / Q000737:chemistry / Q000302:isolation & purification*; D007150:Immunohistochemistry; D051379:Mice; D008856:Microscopy, Fluorescence; D009474:Neurons / Q000737:chemistry* / Q000648:ultrastructure; D009490:Neurosecretory Systems / Q000502:physiology*; D051381:Rats; D016628:Replica Techniques; D013572:Synaptic Vesicles / Q000737:chemistry*
D019204:GTP-Binding Proteins