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Identification and function of conformational dynamics in the multidomain GTP ase dynamin
Author(s) -
Srinivasan Saipraveen,
Dharmarajan Venkatasubramanian,
Reed Dana Kim,
Griffin Patrick R,
Schmid Sandra L
Publication year - 2016
Publication title -
the embo journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.484
H-Index - 392
eISSN - 1460-2075
pISSN - 0261-4189
DOI - 10.15252/embj.201593477
Subject(s) - biology , dynamin , function (biology) , identification (biology) , gtp' , dynamics (music) , biophysics , computational biology , microbiology and biotechnology , biochemistry , physics , receptor , botany , endocytosis , acoustics , enzyme
Vesicle release upon endocytosis requires membrane fission, catalyzed by the large GTP ase dynamin. Dynamin contains five domains that together orchestrate its mechanochemical activity. Hydrogen–deuterium exchange coupled with mass spectrometry revealed global nucleotide‐ and membrane‐binding‐dependent conformational changes, as well as the existence of an allosteric relay element in the α2 S helix of the dynamin stalk domain. As predicted from structural studies, FRET analyses detect large movements of the pleckstrin homology domain ( PHD ) from a ‘closed’ conformation docked near the stalk to an ‘open’ conformation able to interact with membranes. We engineered dynamin constructs locked in either the closed or open state by chemical cross‐linking or deletion mutagenesis and showed that PHD movements function as a conformational switch to regulate dynamin self‐assembly, membrane binding, and fission. This PHD conformational switch is impaired by a centronuclear myopathy‐causing disease mutation, S619L, highlighting the physiological significance of its role in regulating dynamin function. Together, these data provide new insight into coordinated conformational changes that regulate dynamin function and couple membrane binding, oligomerization, and GTP ase activity during dynamin‐catalyzed membrane fission.