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Modulation of Amyloidogenic Protein Self-Assembly Using Tethered Small Molecules

preprint
submitted on 25.09.2020 and posted on 28.09.2020 by Emma Cawood, Nicolas Guthertz, Jessica Ebo, Theodoros Karamanos, Sheena E. Radford FRS, Andrew Wilson

Protein-protein interactions (PPIs) are involved in many of life’s essential biological functions yet are also an underlying cause of several human diseases, including amyloidosis. The modulation of PPIs presents opportunities to gain mechanistic insights into amyloid assembly, particularly through the use of methods which can trap specific intermediates for detailed study. Such information can also provide a starting point for drug discovery. Here, we demonstrate that covalently tethered small molecule fragments can be used to stabilize specific oligomers during amyloid fibril formation, facilitating the structural characterization of these assembly intermediates. We exemplify the power of covalent tethering using the naturally occurring truncated variant (ΔN6) of the human protein β2-microglobulin (β2m), which assembles into amyloid fibrils associated with dialysis-related amyloidosis. Using this approach, we have trapped tetramers formed by ΔN6 under conditions which would normally lead to fibril formation and found that the degree of tetramer stabilization depends on the site of the covalent tether and the nature of the protein-fragment interaction. The covalent protein-ligand linkage enabled structural characterization of these trapped oligomeric species using X-ray crystallography and NMR, providing insight into why tetramer stabilization inhibits amyloid assembly. Our findings highlight the power of “post-translational chemical modification" as a tool to study biological molecular mechanisms.


Funding

Wellcome Trust 109154/Z/15/A

Wellcome Trust 204963

Wellcome Trust 109984

Wellcome Trust 097827/Z/11/A

Wellcome Trust WT094232MA

Wellcome Trust 094232/Z/10/Z

Wellcome Trust 108466/Z/15/Z

EPSRC EP/N035267/1

EPSRC EP/KO39292/1

EPSRC EP/N013573/1

BBSRC BB/M011151/1

ERC 322408

Royal Society Leverhulme Trust Senior Fellowship - SRF/R1/191087

History

Email Address of Submitting Author

a.j.wilson@leeds.ac.uk

Institution

University of Leeds

Country

United Kingdom

ORCID For Submitting Author

0000-0001-9852-6366

Declaration of Conflict of Interest

The authors declare no competing financial interest

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