Total chemical synthesis and biophysical characterization of the minimal isoform of the KChIP2 potassium channel regulatory subunit.
Total chemical synthesis and biophysical characterization of the minimal isoform of the KChIP2 potassium channel regulatory subunit.
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KChIP2 钾通道调节亚基最小亚型的全化学合成和生物物理表征。
DOI：
10.1110/ps.072876107
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发表时间：
2007
期刊：
影响因子：
--
通讯作者：
Kent,StephenBH
中科院分区：
文献类型：
--
作者：
Rajagopal,Sudarshan;Kent,StephenBH
钾通道辅助亚基KChIP 2与心肌细胞中的Kv4.2通道相关，并参与动作电位复极早期的瞬时外向电流（Ito）的调节。作为第一步，生物药理学探测KChIP 2的机制，我们已经化学合成了它的最小的异构体，KChIP 2d，使用Boc化学固相肽合成结合天然化学连接。合成的KChIP 2d蛋白主要是α-螺旋的预测，并成为更多的结构后，结合钙通过1H-NMR和CD光谱评估。合成的KChIP 2d在溶液中处于单体-二聚体平衡，并且有证据表明Kv4.2的N末端肽上有两个单体结合位点。计划未来的研究包括将荧光和自旋标记的探针在KChIP 2d产生的结构信息与电生理研究平行，以阐明KChIP 2d的作用机制。
The potassium channel accessory subunit KChIP2 associates with Kv4.2 channels in the cardiac myocyte and is involved in the regulation of the transient outward current (Ito) during the early phase of repolarization of the action potential. As a first step to biophysically probe the mechanism of KChIP2, we have chemically synthesized its minimal isoform, KChIP2d, using Boc chemistry solid phase peptide synthesis in conjunction with native chemical ligation. The synthetic KChIP2d protein is primarily alpha‐helical as predicted and becomes more structured upon binding calcium as assessed by1H‐NMR and CD spectroscopy. Synthetic KChIP2d is in a monomer‐dimer equilibrium in solution, and there is evidence for two monomer binding sites on an N‐terminal peptide of Kv4.2. Planned future studies include the incorporation of fluorescent and spin labeled probes in KChIP2d to yield structural information in parallel with electrophysiologic studies to elucidate KChIP2d's mechanism of action.