Improving coiled-coil stability by optimizing ionic interactions
Improving coiled-coil stability by optimizing ionic interactions
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DOI：
10.1016/s0022-2836(02)00114-6
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发表时间：
2002-05-03
影响因子：
5.6
通讯作者：
Lustig, A
中科院分区：
文献类型：
--
作者：
Burkhard, P;Ivaninskii, S;Lustig, A
α-螺旋卷曲螺旋是一种常见的蛋白质寡聚化基序，主要通过沿着卷曲螺旋界面发生的疏水相互作用来稳定。我们最近设计并解决了一个两个七肽重复卷曲螺旋肽的结构，该结构通过除了疏水相互作用之外的螺旋间和螺旋内盐桥的复杂网络进一步稳定。在这里，我们扩展和改进的从头设计这两个七重复肽的四个新设计的肽，其特征在于不同类型的离子相互作用。这些不同类型的离子相互作用的卷曲螺旋的稳定性的贡献进行了分析，通过CD光谱和分析超离心。我们发现，所有的肽是高度α-螺旋，其中两个是100%的二聚体在生理条件下。此外，我们还求解了这些肽中最稳定的肽的X射线结构，并通过将该结构与母体肽的结构进行比较来验证合理的设计原则。我们表明，通过结合最有利的螺旋间和螺旋内盐桥的安排，它是可能的设计卷曲螺旋寡聚化域具有改善的稳定性。(C)2002爱思唯尔科技有限公司版权所有。
alpha-Helical coiled coils are a common protein oligomerization motif stabilized mainly by hydrophobic interactions occurring along the coiled-coil interface. We have recently designed and solved the structure of a two-heptad repeat coiled-coil peptide that is stabilized further by a complex network of inter- and intrahelical salt-bridges in addition to the hydrophobic interactions. Here, we extend and improve the de novo design of this two heptad-repeat peptide by four newly designed peptides characterized by different types of ionic interactions. The contribution of these different types of ionic interactions to coiled-coil stability are analyzed by CD spectroscopy and analytical ultracentrifugation. We show that all peptides are highly alpha-helical and two of them are 100% dimeric under physiological conditions. Furthermore, we have solved the X-ray structure of the most stable of these peptides and the rational design principles are verified by comparing this structure to the structure of the parent peptide. We show that by combining the most favorable inter- and intrahelical salt-bridge arrangements it is possible to design coiled-coil oligomerization domains with improved stability properties. (C) 2002 Elsevier Science Ltd. All rights reserved.