Regulation of reversible binding of smg p25A, a ras p21-like GTP-binding protein, to synaptic plasma membranes and vesicles by its specific regulatory protein, GDP dissociation inhibitor.
We have previously purified from bovine brain cytosol a novel regulatory protein for smg p25A, a ras p21-like GTP-binding protein. This protein, named smg p25A GDP dissociation inhibitor (GDI), regulates the GDP/GTP exchange reaction of smg p25A by inhibiting the dissociation of GDP from and thereby the subsequent binding of GTP to it. We have also previously found that smg p25A is mainly localized in presynaptic plasma membranes and vesicles and moderately in presynaptic cytosol in rat brain synapses. In this paper, we have studied the possible involvement of smg p25A GDI in the localization of smg p25A in the cytosol, plasma membranes, and vesicles in rat brain synapses. Both the GDP- and GTP-bound forms of smg p25A bound to the synaptic membranes and vesicles. smg p25A GDI inhibited the binding of the GDP-bound form of smg p25A, but not that of the GTP-bound form, to the synaptic membranes and vesicles. Moreover, smg p25A GDI induced the dissociation of the GDP-bound form, but not that of the GTP-bound form, of both endogenous and exogenous smg p25As from the synaptic membranes and vesicles. smg p25A GDI made a complex with the GDP-bound form of smg p25A with a molar ratio of 1:1, but not with the GTP-bound or guanine nucleotide-free form. These results suggest that smg p25A reversibly binds to synaptic plasma membranes and vesicles and that this reversible binding is regulated by its specific GDI.
Araki
S
S
Department of Biochemistry, Kobe University School of Medicine, Japan.
Kikuchi
A
A
Hata
Y
Y
Isomura
M
M
Takai
Y
Y
D000818:Animals; D001921:Brain / Q000378:metabolism; D002352:Carrier Proteins / Q000302:isolation & purification / Q000378:metabolism*; D002417:Cattle; D003600:Cytosol / Q000378:metabolism; D004910:Erythrocyte Membrane / Q000378:metabolism; D019204:GTP-Binding Proteins / Q000302:isolation & purification / Q000378:metabolism*; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D006153:Guanosine Diphosphate / Q000378:metabolism; D006160:Guanosine Triphosphate / Q000031:analogs & derivatives / Q000378:metabolism; D006706:Homeostasis; D006801:Humans; D007700:Kinetics; D008928:Mitochondria / Q000378:metabolism; D011485:Protein Binding; D011518:Proto-Oncogene Proteins / Q000378:metabolism; D016283:Proto-Oncogene Proteins p21(ras); D013570:Synaptic Membranes / Q000378:metabolism*; D013572:Synaptic Vesicles / Q000378:metabolism*; D013873:Thionucleotides / Q000378:metabolism; D020692:rab3 GTP-Binding Proteins
D002352:Carrier Proteins; D011518:Proto-Oncogene Proteins; D013873:Thionucleotides; D006153:Guanosine Diphosphate; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D006160:Guanosine Triphosphate; D019204:GTP-Binding Proteins; C501009:HRAS protein, human; D016283:Proto-Oncogene Proteins p21(ras); D020692:rab3 GTP-Binding Proteins