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Dimers of D76N-β2-microglobulin display potent anti-amyloid aggregation activity
Maya-Martinez, Roberto and Xu, Yong and Guthertz, Nicolas and Walko, Martin and Sobott, Frank and Karamanos, Theodoros and Breeze, Alex L. and Radford, Sheena E. (2022) Dimers of D76N-β2-microglobulin display potent anti-amyloid aggregation activity. University of Leeds. [Dataset] https://doi.org/10.5518/1159
Dataset description
Self-association of wild-type β2-microglobulin (WT-β2m) into amyloid fibrils is associated with the disorder Dialysis Related Amyloidosis (DRA). In the familial variant D76N-β2m, the single amino acid substitution enhances the aggregation propensity of the protein dramatically and gives rise to a disorder that is independent of renal dysfunction. Numerous biophysical and structural studies on WT- and D76N-β2m have been performed in order to better understand the structure and dynamics of the native proteins and their different potentials to aggregate into amyloid. However, the structural properties of transient D76N-β2m oligomers and their role(s) in assembly remained uncharted. Here we have utilized NMR methods, combined with photo-induced cross-linking, to detect, trap, and structurally characterize transient dimers of D76N-β2m. We show that the cross-linked D76N-β2m dimers have different structures from those previously characterized for the on-pathway dimers of ΔN6-β2m and are unable to assemble into amyloid. Instead, the cross-linked D76N-β2m dimers are potent inhibitors of amyloid formation, preventing primary nucleation and elongation/secondary nucleation when added in sub-stoichiometric amounts with D76N-β2m monomers. The results highlight the specificity of early protein-protein interactions in amyloid formation and show how mapping these interfaces can inform new strategies to inhibit amyloid assembly.
| Keywords: | Amyloidosis, β2-microglobulin, NMR, Crosslinking |
|---|---|
| Divisions: | Faculty of Biological Sciences > Astbury Centre for Structural Molecular Biology |
| License: | Creative Commons Attribution 4.0 International (CC BY 4.0) |
| Date deposited: | 31 Oct 2022 10:05 |
| URI: | http://archive.researchdata.leeds.ac.uk/id/eprint/1040 |
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Documentation
README_notes.docx [14kB]  |
README_notes.docx [14kB] 
README_notes.docx [14kB] Data
De-novo fibrillation of D76N and WT [11kB] |
Intermolecular-PRE-NMR data of S33C, S57C and S88C [4kB] |
Size Exclusion Chromatography profile of XL sample [747kB] |
Restraints used to generate the D76N dimeric model [14kB] |
Fibrillation of D76N in presence of 10% of XL-Dimers [11kB] |
Fibrillation of D76N in presence of 0, 3 and 10% of seeds and supplemented with 10% of XL-Dimers [87kB] |
Melting temperature of D76N-Cys-MTSL-B2m variants [122B] |
Secondary structure of D76N-Cys-MTSL variants [3kB] |
HN-CSP on D76N as result of Cys-MTSL modification [3kB] |
Inter-PRE-NMR data of S20C [1kB] |
Aggregation kinetics of D76N in presence of different percentages of XL-Monomer and Dimers [121kB] |
Pelleting_assay_between_fibrils and XL-D [73B] |
SPR_sensograms of D76N in different concentrations of XL-Dimers [63kB] |