Fragment-linking peptide design yields a high-affinity ligand for microtubule-based transport
Fragment-linking peptide design yields a high-affinity ligand for microtubule-based transport
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DOI：
10.1016/j.chembiol.2021.03.010
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发表时间：
2021-09-16
影响因子：
8.6
通讯作者：
Dodding, Mark P.
中科院分区：
文献类型：
--
作者：
Cross, Jessica A.;Chegkazi, Magda S.;Dodding, Mark P.
合成肽是操纵细胞内蛋白质-蛋白质相互作用的有吸引力的候选者，因为它们模拟天然相互作用以竞争结合。然而，蛋白质-肽相互作用通常是动态的和弱的。一个挑战是设计肽，使改善与目标的相互作用。在这里，我们设计了一个片段连接策略-“混搭”设计-提供一个高亲和力的配体，KinTag，驱动蛋白-1电机。利用天然微摩尔亲和力货物接头配体的结构见解，我们已经确定并结合了单个高亲和力配体的关键结合特征。X-射线晶体结构显示了设计的相互作用，并显示界面面积仅适度增加。此外，当基因编码时，KinTag以比天然序列更高的效率促进溶酶体的运输，揭示了马达-适配器结合亲和力和细胞器运输之间的直接联系。总之，这些数据证明了肽设计的片段连接策略及其在合成马达配体中的应用，以指导细胞货物运输。
Synthetic peptides are attractive candidates to manipulate protein-protein interactions inside the cell as they mimic natural interactions to compete for binding. However, protein-peptide interactions are often dynamic and weak. A challenge is to design peptides that make improved interactions with the target. Here, we devise a fragment-linking strategy-"mash-up'' design-to deliver a high-affinity ligand, KinTag, for the kinesin-1 motor. Using structural insights from natural micromolar-affinity cargo-adaptor ligands, we have identified and combined key binding features in a single, high-affinity ligand. An X-ray crystal structure demonstrates interactions as designed and reveals only a modest increase in interface area. Moreover, when genetically encoded, KinTag promotes transport of lysosomes with higher efficiency than natural sequences, revealing a direct link between motor-adaptor binding affinity and organelle transport. Together, these data demonstrate a fragment-linking strategy for peptide design and its application in a synthetic motor ligand to direct cellular cargo transport.