Affinity-guided protein conjugation: the trilogy of covalent protein labeling, assembly and inhibition
Affinity-guided protein conjugation: the trilogy of covalent protein labeling, assembly and inhibition
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DOI：
10.1007/s11426-016-5571-6
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发表时间：
2016-03
期刊：
影响因子：
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通讯作者：
Yongsheng Yu;Jiang Xia
中科院分区：
文献类型：
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作者：
Yongsheng Yu;Jiang Xia
特定且动态的生物相互作用为细胞内信号网络的蓝图奠定了基础。例如，多种特定的蛋白质-配体相互作用定义了细胞内信号的流动方式。利用这些相互作用的特异性，我们提出了一种“亲和力引导的共价缀合”策略，通过配体和受体蛋白之间的共价反应来锁定结合配体。靠近蛋白质配体结合位点的亲核试剂的存在不是该反应的必要条件。配体衍生物（在设计位置含有亲电子试剂）与蛋白质的配体结合位点的特异性非共价相互作用使亲电子试剂与亲核试剂非常接近。随后，亲核试剂和亲电子试剂之间自发发生共轭反应，并导致分子间共价连接。该策略首先在卷曲螺旋肽中展示，该肽在选定位置包含半胱氨酸突变。短肽序列用于细胞表面受体的共价标记。然后使用相同的策略来指导一组蛋白质乐高积木的设计，用于非自然几何形状的蛋白质复合物的共价组装。我们最终为天然衔接蛋白制造了“反应肽”，在信号转导中发挥重要作用。这些肽被设计为与多结构域衔接蛋白的单个结构域发生反应，被递送到神经元的胞质中，并重新定向神经元迁移的细胞内信号。蛋白质标记、组装和抑制细胞内信号的三部曲，全部通过特定的共价键，充分证明了“亲和引导的共价缀合”在各种应用中的通用性和多功能性。
Specific and dynamic biological interactions pave the blueprint of signal networks in cell. For example, a great variety of specific protein-ligand interactions define how intracellular signals flow. Taking advantage of the specificity of these interactions, we postulate an “affinity-guided covalent conjugation” strategy to lock binding ligands through covalent reactions between the ligand and the receptor protein. The presence of a nucleophile close to the ligand binding site of a protein issine qua noneof this reaction. Specific noncovalent interaction of a ligand derivative (which contains an electrophile at a designed position) to the ligand binding site of the protein brings the electrophile to the close proximity of the nucleophile. Subsequently, a conjugation reaction spontaneously takes place between the nucleophile and the electrophile, and leads to an intermolecular covalent linkage. This strategy was first showcased in coiled coil peptides which include a cysteine mutation at a selected position. The short peptide sequence was used for covalent labeling of cell surface receptors. The same strategy was then used to guide the design of a set of protein Lego bricks for covalent assembly of protein complexes of unnatural geometry. We finally made “reactive peptides” for natural adaptor proteins that play significant roles in signal transduction. The peptides were designed to react with a single domain of the multidomain adaptor protein, delivered into the cytosol of neurons, and re-directed the intracellular signal of neuronal migration. The trilogy of protein labeling, assembly, and inhibition of intracellular signals, all through a specific covalent bond, fully demonstrated the generality and versatility of “affinity-guided covalent conjugation” in various applications.