@article{26924a3f9142400bbc046fbc4c8fdf02,
title = "G domain dimerization controls dynamin's assembly-stimulated GTPase activity",
abstract = "Dynamin is an atypical GTPase that catalyses membrane fission during clathrin-mediated endocytosis. The mechanisms of dynamins basal and assembly-stimulated GTP hydrolysis are unknown, though both are indirectly influenced by the GTPase effector domain (GED). Here we present the 2.0 {\AA} resolution crystal structure of a human dynamin 1-derived minimal GTPase-GED fusion protein, which was dimeric in the presence of the transition state mimic GDP.AlF4-.The structure reveals dynamins catalytic machinery and explains how assembly-stimulated GTP hydrolysis is achieved through G domain dimerization. A sodium ion present in the active site suggests that dynamin uses a cation to compensate for the developing negative charge in the transition state in the absence of an arginine finger. Structural comparison to the rat dynamin G domain reveals key conformational changes that promote G domain dimerization and stimulated hydrolysis. The structure of the GTPase-GED fusion protein dimer provides insight into the mechanisms underlying dynamin-catalysed membrane fission.",
author = "Chappie, {Joshua S.} and Sharmistha Acharya and Marilyn Leonard and Schmid, {Sandra L.} and Fred Dyda",
note = "Funding Information: Acknowledgements We thank V. Lukiyanchuk for assistance in cloning and purification; A. Hickman, J. Mindell and R. Ramachandran for discussions and technical advice; T. Pucadyil and R. Ramachandran for critical reading of the manuscript; J. Hinshaw and J. Mears for providing the unpublished coordinates for the GTPase docking model derived from their cryo-EM docking studies; and R. Stevens and I. Wilson for structural advice, guidance and the use of their laboratory facilities in the early stages of this work. This work was supported by NIH grants GM42455 and MH61345 (to S.L.S.) and the Intramural Program of the National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK) of the NIH. J.S.C. was supported by a Ruth Kirschstein individual predoctoral fellowship from the NIMH (MH081419) and by a postdoctoral Intramural Research Training Award from NIDDK. Data were collected at the SER-CAT 22-ID beamline at the Advanced Photon Source, Argonne National Laboratory. Use of the APS was supported by the US Department of Energy, Basic Energy Sciences, Office of Science, under contract no. W-31-109-Eng-38.",
year = "2010",
month = may,
day = "27",
doi = "10.1038/nature09032",
language = "English (US)",
volume = "465",
pages = "435--440",
journal = "Nature",
issn = "0028-0836",
publisher = "Nature Publishing Group",
number = "7297",
}