Enhancement of cell type specificity by quantitative modulation of a chimeric ligand
Enhancement of cell type specificity by quantitative modulation of a chimeric ligand
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DOI：
10.1074/jbc.m708502200
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发表时间：
2008-03-28
影响因子：
4.8
通讯作者：
Silver, Pamela A.
中科院分区：
文献类型：
--
作者：
Cironi, Pablo;Swinburne, Ian A.;Silver, Pamela A.
进化调节了蛋白质相互作用的数量特征，并经常使用弱相互作用的组合来达到特定的特异性。我们讨论了如何在多结构域蛋白的设计中使用定量优化，使用含有表皮生长因子（EGF）作为细胞靶向元件和干扰素α -2 α (IFN α -2 α)的嵌合体来启动信号转导。我们首先通过一个连接体连接EGF和IFN α -2 α，该连接体允许两种配体在细胞表面与它们的受体结合，然后将一系列突变纳入IFN α -2 α部分，这些突变逐渐降低了IFN α -2 α -IFN α α受体2 （IFNAR2）相互作用的开启率和解离常数。使用这种策略，我们设计了嵌合蛋白，其中HeLa， A431和工程Daudi细胞中的IFN α受体的激活依赖于同一细胞上EGF受体的存在。突变嵌合蛋白也以EGF受体依赖的方式抑制IFN α敏感细胞的增殖。这些结果提供了对多亚基受体特异性结合的定量要求的见解，并说明了定量方法在合成生物学结构设计中的价值。
Evolution modulates the quantitative characteristics of protein interactions and often uses combinations of weak interactions to achieve a particular specificity. We addressed how quantitative optimization might be used in the design of multidomain proteins, using a chimera containing epidermal growth factor (EGF) as a cell targeting element and interferon alpha-2 alpha (IFN alpha-2 alpha) to initiate signal transduction. We first connected EGF and IFN alpha-2 alpha via a linker that allows both ligands to bind to their receptors on a cell surface and then incorporated a series of mutations into the IFN alpha-2 alpha portion that progressively decrease both the on rate and the dissociation constant of the IFN alpha-2 alpha-IFN alpha receptor 2 (IFNAR2) interaction. Using this strategy, we designed chimeric proteins in which the activation of the IFN alpha receptor in HeLa, A431, and engineered Daudi cells depends on the presence of EGF receptor on the same cell. The mutant chimeric proteins also inhibited proliferation of IFN alpha-sensitive cells in an EGF receptor-dependent manner. These results provide insights into the quantitative requirements for specific binding to multisubunit receptors and illustrate the value of a quantitative approach in the design of synthetic-biological constructs.