Construction of a Ca2+-gated Artificial Channel by Fusing Alamethicin with a Calmodulin-derived Extramembrane Segment
Construction of a Ca2+-gated Artificial Channel by Fusing Alamethicin with a Calmodulin-derived Extramembrane Segment
复制标题
通过将阿拉美辛与钙调蛋白衍生的膜外片段融合构建 Ca2 门控人工通道
DOI：
10.1021/bc300468x
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发表时间：
2013
期刊：
影响因子：
--
通讯作者：
Shiroh Futaki
中科院分区：
文献类型：
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作者：
Daisuke Noshiro;Kazuhiro Sonomura;Hao-Hsin Yu;Miki Imanishi;Koji Asami;Shiroh Futaki
使用天然化学连接，我们构建了一个钙门控融合通道蛋白丙甲霉素和钙调素的C-末端结构域。在pH 5.4和在Ca 2+的情况下，这种融合蛋白产生了一个突发式的通道电流，没有离散的通道电导水平。然而，Ca ~（2+）显著延长了特定通道开放状态，并增加了平均通道电流，而Mg ~（2+）对通道电流无显著影响。8-苯胺基萘-1-磺酸（ANS）荧光测量的基础上，钙离子刺激门控可能与增加的融合蛋白的膜外段的表面疏水性。
Using native chemical ligation, we constructed a Ca2+-gated fusion channel protein consisting of alamethicin and the C-terminal domain of calmodulin. At pH 5.4 and in the absence of Ca2+, this fusion protein yielded a burst-like channel current with no discrete channel conductance levels. However, Ca2+significantly lengthened the specific channel open state and increased the mean channel current, while Mg2+produced no significant changes in the channel current. On the basis of 8-anilinonaphthalene-1-sulfonic acid (ANS) fluorescent measurement, Ca2+-stimulated gating may be related to an increased surface hydrophobicity of the extramembrane segment of the fusion protein.