J Am Chem Soc. 2020 Mar 11;142(10):4534-4538. doi: 10.1021/jacs.9b12003. Epub 2020 Mar 3.
This paper presents a solid-phase strategy to efficiently assemble multiprotein scaffolds-known as megamolecules-without the need for protecting groups and with precisely defined nanoscale architectures. The megamolecules are assembled through sequential reactions of linkers that present irreversible inhibitors for enzymes and fusion proteins containing the enzyme domains. Here, a fusion protein containing an N-terminal cutinase and a C-terminal SnapTag domain react with an ethyl -nitrophenyl phosphonate (pNPP) or a chloro-pyrimidine (CP) group, respectively, to give covalent products. By starting with resin beads that are functionalized with benzylguanine, a series of reactions lead to linear, branched, and dendritic structures that are released from the solid support by addition of TEV protease and that have sizes up to approximately 25 nm.
本文提出了一种固相策略，可有效地组装多蛋白支架——即巨型分子，而无需保护基团，并具有精确定义的纳米级结构。通过含有酶域的连接子的顺序反应来组装巨型分子，这些连接子为酶和融合蛋白提供不可逆的抑制剂。在这里，一种含有 N 端角质酶和 C 端 SnapTag 结构域的融合蛋白分别与乙基-硝基苯膦酸酯（pNPP）或氯嘧啶（CP）基团反应，得到共价产物。通过起始于功能化的苯甲基鸟嘌呤的树脂珠，一系列反应导致线性、支化和树突状结构，通过添加 TEV 蛋白酶从固体载体上释放出来，并且大小可达约 25nm。
