nmr derived model of gtpase effector domain (ged) self association relevance to dynamin assemblynmr派生gtpase效应模型域(ged)自我协会相关dynamin组装.pdfVIP

NMR Derived Model of GTPase Effector Domain (GED) Self Association: Relevance to Dynamin Assembly Swagata Chakraborty, Supriya Pratihar, Ramakrishna V. Hosur* Department of Chemical Sciences, Tata Institute of Fundamental Research, Mumbai, India Abstract Self-association of dynamin to form spiral structures around lipidic vesicles during endocytosis is largely mediated by its ‘coiled coil’ GTPase Effector Domain (GED), which, in vitro, self-associates into huge helical assemblies. Residue-level structural characterizations of these assemblies and understanding the process of association have remained a challenge. It is also impossible to get folded monomers in the solution phase. In this context, we have developed here a strategy to probe the self-association of GED by first dissociating the assembly using Dimethyl Sulfoxide (DMSO) and then systematically monitoring the refolding into helix and concomitant re-association using NMR spectroscopy, as DMSO concentration is progressively reduced. The short segment, Arg109 - Met116, acts as the nucleation site for helix formation and self-association. Hydrophobic and complementary charge interactions on the surfaces drive self-association, as the helices elongate in both the directions resulting in an antiparallel stack. A small N-terminal segment remains floppy in the assembly. Following these and other published results on inter-domain interactions, we have proposed a plausible mode of dynamin self assembly. Citation: Chakraborty S, Pratihar S, Hosur RV (2012) NMR Derived Model of GTPase Effector Domain (GED) Self Association: Relevance to Dynamin Assembly. PLoS ONE 7(1): e30109. doi:10.1371/journal.pone.0030109 Editor: Maria Gasset, Consejo Superior de Investigaciones Cientificas, Spain Received September 16, 2011; Accepted December