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TitleD-loop Dynamics and Near-Atomic-Resolution Cryo-EM Structure of Phalloidin-Bound F-Actin.
Journal, issue, pagesStructure, Vol. 28, Issue 5, Page 586-593.e3, Year 2020
Publish dateMay 5, 2020
AuthorsSanchaita Das / Peng Ge / Zeynep A Oztug Durer / Elena E Grintsevich / Z Hong Zhou / Emil Reisler /
PubMed AbstractDetailed molecular information on G-actin assembly into filaments (F-actin), and their structure, dynamics, and interactions, is essential for understanding their cellular functions. Previous studies ...Detailed molecular information on G-actin assembly into filaments (F-actin), and their structure, dynamics, and interactions, is essential for understanding their cellular functions. Previous studies indicate that a flexible DNase I binding loop (D-loop, residues 40-50) plays a major role in actin's conformational dynamics. Phalloidin, a "gold standard" for actin filament staining, stabilizes them and affects the D-loop. Using disulfide crosslinking in yeast actin D-loop mutant Q41C/V45C, light-scattering measurements, and cryoelectron microscopy reconstructions, we probed the constraints of D-loop dynamics and its contribution to F-actin formation/stability. Our data support a model of residues 41-45 distances that facilitate G- to F-actin transition. We report also a 3.3-Å resolution structure of phalloidin-bound F-actin in the ADP-Pi-like (ADP-BeFx) state. This shows the phalloidin-binding site on F-actin and how the relative movement between its two protofilaments is restricted by it. Together, our results provide molecular details of F-actin structure and D-loop dynamics.
External linksStructure / PubMed:32348747 / PubMed Central
MethodsEM (helical sym.)
Resolution3.8 Å
Structure data

20694

6u96

EMDB-20694, PDB-6u96:
Actin phalloidin at BeFx state
Method: EM (helical sym.) / Resolution: 3.8 Å

Chemicals

ChemComp-ADP:
ADENOSINE-5'-DIPHOSPHATE / ADP, energy-carrying molecule*YM

Source
  • oryctolagus cuniculus (rabbit)
  • Rabbit (rabbit)
  • synthetic construct (others)
KeywordsSTRUCTURAL PROTEIN / Actin / phalloidin / beryllium fluoride

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