Chugh Jeetender, Sharma Shilpy, Hosur Ramakrishna V
Department of Chemical Sciences, Tata Institute of Fundamental Research, Mumbai 400005, India.
Protein Sci. 2008 Aug;17(8):1319-25. doi: 10.1110/ps.035840.108. Epub 2008 May 27.
Protein self-association is critical to many biological functions. However, atomic-level structural characterization of these assemblies has remained elusive. In this report we present insights into the mechanistic details of the process of self-association of the 136-residue GTPase effector domain (GED) of the endocytic protein dynamin into a megadalton-sized soluble mass. Our approach is based on NMR monitoring of regulated folding and association through Gdn-HCl titration. The results suggest the evolution of a sequence-self-association paradigm. Equally significantly, the study demonstrates an elegant bottom-up strategy that can render large protein self-assemblies accessible to NMR investigations that have remained difficult to date.
蛋白质自组装对许多生物学功能至关重要。然而，这些组装体的原子水平结构表征仍然难以捉摸。在本报告中，我们深入探讨了内吞蛋白发动蛋白的136个残基的GTPase效应结构域（GED）自组装成兆道尔顿大小的可溶性聚集体过程的机制细节。我们的方法基于通过盐酸胍滴定对内源折叠和组装进行核磁共振监测。结果表明了一种序列-自组装模式的演变。同样重要的是，该研究展示了一种精妙的自下而上的策略，该策略可以使大型蛋白质自组装体能够进行核磁共振研究，而迄今为止这一直是困难的。
