Ichida S, Wada T, Akimoto T, Kasamatsu Y, Tahara M, Hasimoto K
Department of Biological Chemistry, Faculty of Pharmacy, Kinki University, Higashi-Osaka, Japan.
Neurochem Res. 1995 Apr;20(4):467-73. doi: 10.1007/BF00973104.
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.
使用双功能交联剂（二硫代双[琥珀酰亚胺丙酸酯]：DSP）对鸡全脑粗膜中[125I]ω-芋螺毒素（ω-CgTX）标记的特性进行了系统研究。在非还原条件下，[125I]ω-CgTX在粗膜中特异性标记出一条216 kDa的主要条带和一条236 kDa的次要条带，但在还原条件下未标记。我们详细研究了各种钙通道拮抗剂对用DSP进行的[125I]ω-CgTX标记的影响，发现钙通道拮抗剂对216 kDa条带的[125I]ω-CgTX标记的影响与[125I]ω-CgTX的特异性结合之间存在很强的相关性。这些结果表明，在非还原条件下使用DSP用[125I]ω-CgTX对216 kDa条带进行标记涉及[125I]ω-CgTX的特异性结合位点，可能包括粗膜中神经元N型钙通道亚基之一。
