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Elife. 2018 Jan 23;7. pii: e30246. doi: 10.7554/eLife.30246.

Dynamic clustering of dynamin-amphiphysin helices regulates membrane constriction and fission coupled with GTP hydrolysis.

Author information

1
Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, Okayama, Japan.
2
Department of Physics, College of Science and Engineering, Kanazawa University, Kanazawa, Japan.
3
CREST, JST, Saitama, Japan.
4
Department of Physics, School of Science, Nagoya University, Nagoya, Japan.
5
Bio-AFM Frontier Research Center, College of Science and Engineering, Kanazawa University, Kanazawa, Japan.

Abstract

Dynamin is a mechanochemical GTPase essential for membrane fission during clathrin-mediated endocytosis. Dynamin forms helical complexes at the neck of clathrin-coated pits and their structural changes coupled with GTP hydrolysis drive membrane fission. Dynamin and its binding protein amphiphysin cooperatively regulate membrane remodeling during the fission, but its precise mechanism remains elusive. In this study, we analyzed structural changes of dynamin-amphiphysin complexes during the membrane fission using electron microscopy (EM) and high-speed atomic force microscopy (HS-AFM). Interestingly, HS-AFM analyses show that the dynamin-amphiphysin helices are rearranged to form clusters upon GTP hydrolysis and membrane constriction occurs at protein-uncoated regions flanking the clusters. We also show a novel function of amphiphysin in size control of the clusters to enhance biogenesis of endocytic vesicles. Our approaches using combination of EM and HS-AFM clearly demonstrate new mechanistic insights into the dynamics of dynamin-amphiphysin complexes during membrane fission.

KEYWORDS:

EM; HS-AFM; amphiphysin; biophysics; cell biology; dynamin; human; in vitro reconstitution; membrane remodeling; structural biology

PMID:
29357276
PMCID:
PMC5780043
DOI:
10.7554/eLife.30246
[Indexed for MEDLINE]
Free PMC Article

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