A peptide-display protein scaffold to facilitate single molecule force studies of aggregation-prone peptides

作者：Doherty Ciaran P A; Young Lydia M; Karamanos Theodoros K; Smith Hugh I; Jackson Matthew P; Radford Sheena E; Brockwell David J^*

来源：Protein Science, 2018, 27(7): 1205-1217.

DOI：10.1002/pro.3386

• single molecule force spectroscopy
• alpha-synuclein
• gamma-synuclein
• aggregation
• amyloid
• mass spectrometry

Protein aggregation is linked with the onset of several neurodegenerative disorders, including Parkinson's disease (PD), which is associated with the aggregation of -synuclein (Syn). The structural mechanistic details of protein aggregation, including the nature of the earliest protein-protein interactions, remain elusive. In this study, we have used single molecule force spectroscopy (SMFS) to probe the first dimerization events of the central aggregation-prone region of Syn (residues 71-82) that may initiate aggregation. This region has been shown to be necessary for the aggregation of full length Syn and is capable of forming amyloid fibrils in isolation. We demonstrate that the interaction of Syn(71-82) peptides can be studied using SMFS when inserted into a loop of protein L, a mechanically strong and soluble scaffold protein that acts as a display system for SMFS studies. The corresponding fragment of the homolog protein -synuclein (Syn), which has a lower aggregation propensity, has also been studied here. The results from SMFS, together with native mass spectrometry and aggregation assays, demonstrate that the dimerization propensity of Syn(71-82) is lower than that of Syn(71-82), but that a mixed Syn(71-82): Syn(71-82) dimer forms with a similar propensity to the Syn(71-82) homodimer, slowing amyloid formation. This work demonstrates the utility of a novel display method for SMFS studies of aggregation-prone peptides, which would otherwise be difficult to study.

• 出版日期2018-7