Cooperative amyloid fibre binding and disassembly by the Hsp70 disaggregase

DOI: 10.15252/embj.2021110410

Authors: Joseph G. Beton (Birkbeck University of London) , Jim Monistrol (Birkbeck University of London) , Anne Wentink (Center for Molecular Biology of Heidelberg University (ZMBH) and German Cancer Research Center (DKFZ), DKFZ-ZMBH Alliance) , Erin C. Johnston (Birkbeck University of London) , Anthony J. Roberts (Birkbeck University of London) , Bernd G. Bukau (Center for Molecular Biology of Heidelberg University (ZMBH) and German Cancer Research Center (DKFZ), DKFZ-ZMBH Alliance) , Bart W. Hoogenboom (University College London) , Helen R. Saibil (Birkbeck University of London)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: The Embo Journal , VOL 9

State: Published (Approved)
Published: June 2022
Diamond Proposal Number(s): 20287

Abstract: Although amyloid fibres are highly stable protein aggregates, a specific combination of human Hsp70 system chaperones can disassemble them, including fibres formed of α-synuclein, huntingtin, or Tau. Disaggregation requires the ATPase activity of the constitutively expressed Hsp70 family member, Hsc70, together with the J domain protein DNAJB1 and the nucleotide exchange factor Apg2. Clustering of Hsc70 on the fibrils appears to be necessary for disassembly. Here we use atomic force microscopy to show that segments of in vitro assembled α-synuclein fibrils are first coated with chaperones and then undergo bursts of rapid, unidirectional disassembly. Cryo-electron tomography and total internal reflection fluorescence microscopy reveal fibrils with regions of densely bound chaperones, preferentially at one end of the fibre. Sub-stoichiometric amounts of Apg2 relative to Hsc70 dramatically increase recruitment of Hsc70 to the fibres, creating localised active zones that then undergo rapid disassembly at a rate of ~ 4 subunits per second. The observed unidirectional bursts of Hsc70 loading and unravelling may be explained by differences between the two ends of the polar fibre structure.

Journal Keywords: atomic force microscopy; cryo-electron tomography; disaggregation; molecular chaperones

Subject Areas: Biology and Bio-materials

Diamond Offline Facilities: Electron Bio-Imaging Centre (eBIC)
Instruments: Krios I-Titan Krios I at Diamond

Added On: 19/06/2022 10:27

Documents:
The EMBO Journal - 2022 - Beton - Cooperative amyloid fibre binding and disassembly by the Hsp70 disaggregase.pdf

Discipline Tags:

Neurodegenerative Diseases Non-Communicable Diseases Health & Wellbeing Neurology Structural biology Life Sciences & Biotech

Technical Tags:

Microscopy Electron Microscopy (EM) Cryo Electron Microscopy (Cryo EM)