KChIP3:: A binding protein for Taiwan banded krait β-bungarotoxin
KChIP3:: A binding protein for Taiwan banded krait β-bungarotoxin
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DOI：
10.1016/j.toxicon.2005.10.020
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发表时间：
2006-03-01
期刊：
影响因子：
2.8
通讯作者：
Chang, LS
中科院分区：
文献类型：
--
作者：
Lin, YL;Wu, PF;Chang, LS
以β -bungarotoxin (β - bgt) B1链为诱饵进行酵母双杂交筛选，发现KChIP3是B1链的结合蛋白。因此，在本研究中研究了β - bgt和KChIP3之间的蛋白-蛋白相互作用。Pull-down实验显示重组KChIP3蛋白与β - bgt和B1链相关，而kchip1、2和4蛋白不能与β - bgt结合。虽然Ca2+不是KChIP3与β - bgt和B1链结合的关键因素，但它们的相互作用可以通过Ca2+的加入而增强。另外，β - bgt A1链与KChIP3的关联被轻微检测到。在2 mM Ca2+不存在和存在的情况下，β - bgt与KChIP3的解离常数分别为12.2和6.08 μ M。此外，利用β - bgt - sepharose从大鼠脑中分离天然KChIP3。这些观察结果表明KChIP3是β - bgt的结合蛋白。鉴于KChIP3在神经细胞中的多种功能，KChIP3与β - bgt的相互作用可能是β - bgt生物学活性表现的一个事件。(c) 2005 Elsevier Ltd所有航班预订。
Using B1 chain of beta-bungarotoxin (beta-Bgt) as bait in yeast two-hybrid screen, we found that KChIP3 was a binding protein of B1 chain. Thus, protein-protein interaction between beta-Bgt and KChIP3 is investigated in the present study. Pull-down assay showed that recombinant KChIP3 proteins were associated with beta-Bgt as well as B1 chain, whereas the inability of KChIPs 1, 2 and 4 to bind with beta-Bgt was observed. Although Ca2+ was not a crucial factor essential for the binding of KChIP3 with beta-Bgt and B1 chain, their interaction could be enhanced by the addition of Ca2+. Alternatively, the association of A1 chain of beta-Bgt with KChIP3 was marginally detected. The dissociation constant of beta-Bgt with KChIP3 were 12.2 and 6.08 mu M in the absence and presence of 2 mM Ca2+, respectively. Moreover, native KChIP3 from rat brain was to be isolated by beta-Bgt-Sepharose. These observations indicate that KChIP3 is a binding protein of beta-Bgt. In view of the multiple functions of KChIP3 in neuronal cells, the interaction of KChIP3 with beta-Bgt may represent an event for the manifestation of the biological activities of beta-Bgt. (c) 2005 Elsevier Ltd. All fights reserved.