A synthetic coiled-coil interactome provides heterospecific modules for molecular engineering.

A synthetic coiled-coil interactome provides heterospecific modules for molecular engineering.
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DOI:
10.1021/ja907617a
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发表时间:
2010-05-05
影响因子:
15
通讯作者:
Keating, Amy E.
Keating, Amy E.
中科院分区:
化学1区
文献类型:
--
作者:
Reinke, Aaron W.;Grant, Robert A.;Keating, Amy E.

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多功能卷曲蛋白基序在生物学和材料科学中广泛用于诱导和控制大分子相互作用。然而,可以使用已知的线圈构造的交互模式类型是有限的。在这里,我们通过使用肽微阵列测量48个合成线圈和7个人类bZIP线圈的完全成对相互作用,大大扩展了线圈工具箱。由此产生的55个成员的蛋白质“相互作用组”包括27对优先异结合的相互作用肽。这27对蛋白质可以组合成3到6个蛋白质,组成不同拓扑结构的网络。特别感兴趣的是参与相互正交相互作用的异特异性肽对。这样的对提供了机会,使两个独立的分子系统二聚化,而没有不必要的串扰。两组正交异源二聚体的解和结构表征提供了它们相互作用几何的细节。正交对,以及我们在屏幕中发现的许多其他网络基序,为合成生物学和其他应用提供了新的功能。
The versatile coiled-coil protein motif is widely used to induce and control macromolecular interactions in biology and materials science. Yet the types of interaction patterns that can be constructed using known coiled coils are limited. Here we greatly expand the coiled-coil toolkit by measuring the complete pair-wise interactions of 48 synthetic coiled coils and 7 human bZIP coiled coils using peptide microarrays. The resulting 55-member protein ‘interactome’ includes 27 pairs of interacting peptides that preferentially hetero-associate. The 27 pairs can be used in combinations to assemble sets of 3 to 6 proteins that compose networks of varying topologies. Of special interest are heterospecific peptide pairs that participate in mutually orthogonal interactions. Such pairs provide the opportunity to dimerize two separate molecular systems without undesired crosstalk. Solution and structural characterization of two such sets of orthogonal heterodimers provide details of their interaction geometries. The orthogonal pair, along with the many other network motifs discovered in our screen, provide new capabilities for synthetic biology and other applications.
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