Compounds targeting aggregated alpha-synuclein are high in demand as potential bio-imaging tools. We have previously developed an assay that measures the binding of the protein p25alpha to aggregated alpha-synuclein. The rationale for the present project is to refine this assay as a potential tool to screen for alpha-synuclein aggregate binding compounds and as a proof of concept determine if the aggregate binding dye thioflavine-T can displace the binding of p25alpha.
The assay will be developed in a 96 well format compatible with medium through-put screening. In the plates alpha-synuclein filaments will be immobilized. Next labeled p25alpha protein will be bound to the aggregates in the absence and presence of various competitors, e.g. Thioflaine-T. Finally the binding of labeled p25alpha will be determined and the inhibitory effect of the potential competitors determined.
Relevance to Diagnosis/Treatment of Parkinson’s Disease:
Successful development of the assay may allow it to be used for the screening of novel alpha-synuclein aggregate binding compounds that may be developed into bio-imaging tools. Such tools will be critical for future neuroprotective trials and may also serve as diagnostic tools.
We expect to develop an assay the can be used for future screenings for alpha-synuclein aggregate binding compounds.