Antos John M, Popp Maximilian Wei-Lin, Ernst Robert, Chew Guo-Liang, Spooner Eric, Ploegh Hidde L
Whitehead Institute for Biomedical Research, Massachusetts Institute of Technology, Cambridge, Massachusetts 02142, USA.
J Biol Chem. 2009 Jun 5;284(23):16028-36. doi: 10.1074/jbc.M901752200. Epub 2009 Apr 9.
Folding and stability are parameters that control protein behavior. The possibility of conferring additional stability on proteins has implications for their use in vivo and for their structural analysis in the laboratory. Cyclic polypeptides ranging in size from 14 to 78 amino acids occur naturally and often show enhanced resistance toward denaturation and proteolysis when compared with their linear counterparts. Native chemical ligation and intein-based methods allow production of circular derivatives of larger proteins, resulting in improved stability and refolding properties. Here we show that circular proteins can be made reversibly with excellent efficiency by means of a sortase-catalyzed cyclization reaction, requiring only minimal modification of the protein to be circularized.
折叠和稳定性是控制蛋白质行为的参数。赋予蛋白质额外稳定性的可能性对其在体内的应用及其在实验室中的结构分析具有重要意义。天然存在大小从14到78个氨基酸不等的环状多肽，与线性对应物相比，它们通常对变性和蛋白水解表现出更强的抗性。天然化学连接和基于内含肽的方法可用于生产更大蛋白质的环状衍生物，从而提高稳定性和重折叠特性。在这里，我们表明，通过分选酶催化的环化反应，可以高效可逆地制备环状蛋白质，只需对要环化的蛋白质进行最少的修饰。
