In cellulo library-derived peptide-based inhibitors of alpha-synuclein aggregation and toxicity — ASN Events

In cellulo library-derived peptide-based inhibitors of alpha-synuclein aggregation and toxicity (#21)

Richard Meade 1 , Robert J Williams 1 , Jody Mason 1
  1. University of Bath, Bath, NORTH SOMERSET, United Kingdom

A major group focus is the design and selection of peptides that target amyloidogenic proteins involved in age-related diseases. Amyloid proteins are known to be important in a number of such diseases that include Alzheimer’s, Parkinson’s, Lewy Body Dementia, Huntington’s, and CJD. We use a novel in cellulo library-screening platform to select peptides that can bind amyloidogenic target proteins to sequester and detoxify them. Utimising a Protein-fragment Complementation approach (PCA), we have identified both strand and helix-based peptide antagonists of α-synuclein proteins implicated in PD and related synucleinopathies. PCA is multiplexed, making no mechanistic assumptions about the target oligomeric state or conformer populated. Rather, library members must bind to and reduce associated toxicity to become selected. Those that either generate, or fail to prevent formation of a toxic species, result in cell death or retarded cell growth rates relative to effective binders. Library members that confer the most rapid bacterial growth are then selected from the PCA by increased stringency during further competition selection. Our antagonists have been characterised using a range of biophysical and cell-based approaches and been downsized / refined using truncation, alanine-scanning, and incorporation of structure-inducing constraints and non-natural sequences. Our work in this area is currently funded by an Alzheimer’s Research UK Major Project Award.

  1. Cheruvara H, Allen-Baume V.L, Kad N.M, Mason J.M. Intracellular screening of a peptide library to derive a potent peptide inhibitor of α-synuclein aggregation. J Biol Chem. 290, 7426-35 (2015).
  2. Meade, R., Fairlie, D.P., and Mason, J.M. Alpha-Synuclein Structure and Parkinsons Disease: Lessons and Emerging Principles. Mol Neurodegener 14:29 (2019).
  3. Meade, R.M., Morris, K.J., Watt, K.J.C., Williams, R.J., and Mason. J.M. The Library Derived 4554W Peptide Inhibits Primary Nucleation of α-Synuclein. J. Mol. Biol. 432, 166706 (2020).
  4. Meade, R.M., Williams, R.J. and Mason, J.M. A Series of Helical Alpha-synuclein Fibril Polymorphs Are Populated in the Presence of Lipid Vesicles. Nature Partner Journal - Parkinsons disease 6(20), (2020).
  5. Meade, R.M., Watt, K.J.C., Williams, R.J., and Mason. J.M. A Downsized and Optimised Intracellular Library-derived Peptide Prevents α-Synuclein Primary Nucleation and Toxicity Without Impacting Upon Lipid Binding. J. Mol. Biol. 433:167323 (2021).
  6. Watt K.J.C., Meade R.M., Williams R.J., and Mason J.M. Library-derived peptide aggregation modulators of Parkinson's disease early-onset alpha-synuclein variants. ACS Chem. Neurosci. 13, 12, 1790-1804 (2022).
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