Characteristics of [125I]omega-conotoxin labeling using bifunctional cross linker DSP in crude membranes from chick brain.
Characteristic of [125I]omega-conotoxin (omega-CgTX) labeling using bifunctional cross linker (dithio bis[succinimidyl propionate]:DSP) was systematically investigated in crude membranes from chick whole brain. [125I]omega-CgTX specifically labeled 216 kDa as a main and 236 kDa as a minor bands in the crude membranes under non-reduced condition, but not labeled under reduced condition. We investigated the effect of various Ca channel antagonists on [125I]omega-CgTX labeling with DSP in detail, and found that there is a strong correlation between the effects of Ca channel antagonists on [125I]omega-CgTX labeling of the 216 kDa band and specific [125I]omega-CgTX binding. These results suggest that labeling of the 216 kDa band under non-reduced condition with [125I]omega-CgTX using DSP involves the specific binding sites of [125I]omega-CgTX, perhaps including one of the neuronal N-type Ca channel subunits in the crude membranes.
Ichida
S
S
Department of Biological Chemistry, Faculty of Pharmacy, Kinki University, Higashi-Osaka, Japan.
Wada
T
T
Akimoto
T
T
Kasamatsu
Y
Y
Tahara
M
M
Hasimoto
K
K
D000818:Animals; D001921:Brain / Q000378:metabolism*; D002121:Calcium Channel Blockers / Q000378:metabolism*; D002645:Chickens; D003432:Cross-Linking Reagents; D007457:Iodine Radioisotopes; D008566:Membranes / Q000378:metabolism; D008978:Mollusk Venoms / Q000378:metabolism*; D010455:Peptides / Q000378:metabolism*; D011485:Protein Binding; D013388:Succinimides; D020866:omega-Conotoxin GVIA
D002121:Calcium Channel Blockers; D003432:Cross-Linking Reagents; D007457:Iodine Radioisotopes; D008978:Mollusk Venoms; D010455:Peptides; D013388:Succinimides; D020866:omega-Conotoxin GVIA; C011240:dithiobis(succinimidylpropionate)