Objective/Rationale:
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.
Project Description:
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.
Anticipated Outcome:
We expect to develop an assay the can be used for future screenings for alpha-synuclein aggregate binding compounds.