NMR insights into a megadalton‐size protein self‐assembly

Abstract

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.