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RESEARCH PRODUCT
Uncoupling of dynamin polymerization and GTPase activity revealed by the conformation-specific nanobody dynab
Jason EcardAurélien RouxSandrine MoutelSandrine MoutelFranck PerezValentina GalliRafael Sebastiansubject
0301 basic medicineendocrine systemGTP'MouseQH301-705.5FissionScienceEndocytic cycleGTPasemacromolecular substancesEndocytosisGeneral Biochemistry Genetics and Molecular BiologyGTP PhosphohydrolasesPolymerization03 medical and health sciences0302 clinical medicineMembrane fissiondynaminendocytosisHumansBiology (General)Dynamin IDynaminGeneral Immunology and MicrobiologyChemistryGeneral Neuroscienceconformational-specific nanobodyHydrolysisQRGeneral MedicineCell BiologyFibroblastsSingle-Domain Antibodiesenzyme030104 developmental biologyMembraneddc:540BiophysicsMedicineGuanosine Triphosphatebiological phenomena cell phenomena and immunitycell biology conformational-specific nanobody dynamin endocytosis enzyme human mouse030217 neurology & neurosurgeryResearch ArticleHumandescription
Dynamin is a large GTPase that forms a helical collar at the neck of endocytic pits, and catalyzes membrane fission (Schmid and Frolov, 2011; Ferguson and De Camilli, 2012). Dynamin fission reaction is strictly dependent on GTP hydrolysis, but how fission is mediated is still debated (Antonny et al., 2016): GTP energy could be spent in membrane constriction required for fission, or in disassembly of the dynamin polymer to trigger fission. To follow dynamin GTP hydrolysis at endocytic pits, we generated a conformation-specific nanobody called dynab, that binds preferentially to the GTP hydrolytic state of dynamin-1. Dynab allowed us to follow the GTPase activity of dynamin-1 in real-time. We show that in fibroblasts, dynamin GTP hydrolysis occurs as stochastic bursts, which are randomly distributed relatively to the peak of dynamin assembly. Thus, dynamin disassembly is not coupled to GTPase activity, supporting that the GTP energy is primarily spent in constriction.
| year | journal | country | edition | language |
|---|---|---|---|---|
| 2017-10-01 | eLife |