1. Site-specific fluorescence polarization for studying the disaggregation of α-synuclein fibrils by small molecules
- Author
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Merve Canyurt, Malcolm J. Daniels, Lily Owei, Jaclyn Robustelli, Tobias Baumgart, Christina L. Cleveland, Rebecca F. Wissner, Conor M. Haney, Harry Ischiropoulos, E. James Petersson, and Priscilla Rodriguez
- Subjects
0301 basic medicine ,Protein aggregates ,Dopamine ,Fibril formation ,Plasma protein binding ,Protein aggregation ,Biochemistry ,Micelle ,Catechin ,Nordihydroguaiaretic acid ,Fluorescence polarization ,Polarization ,Dodecyl sulfate sodium ,Polyphenol derivative ,Priority journal ,Recombinant proteins ,Analogs and derivatives ,Chemistry ,Vesicle ,Small molecules ,Sodium Dodecyl Sulfate ,Molecular interaction ,Ddrug effects ,Small molecule ,Recombinant Proteins ,Fibrillar aggregates ,Epigallocatechin gallate ,Liposome ,alpha-Synuclein ,Phosphatidylcholines ,Site-specific ,Human ,Binding sites ,Molecular library ,Fluorescence Polarization ,Structural remodeling ,macromolecular substances ,Fibril ,Unilamellar liposomes ,Article ,Fluorescence ,Small Molecule Libraries ,03 medical and health sciences ,Protein Aggregates ,Disaggregation ,Texas red ,Conformational transition ,Humans ,Protein binding ,Masoprocol ,Amino Acid Sequence ,Binding site ,Sodium dodecyl sulfate ,Unilamellar Liposomes ,Fluorescent Dyes ,Pharmacology ,030102 biochemistry & molecular biology ,Xanthene derivative ,Fluorescent dye ,Proteins ,Molecules ,Crystallography ,030104 developmental biology ,Metabolism ,Xanthenes ,Local dynamics ,Biophysics ,Fluorescence anisotropy - Abstract
Fibrillar aggregates of the protein α-synuclein (αS) are one of the hallmarks of Parkinson’s disease. Here, we show that measuring the fluorescence polarization (FP) of labels at several sites on αS allows one to monitor changes in the local dynamics of the protein after binding to micelles or vesicles, and during fibril formation. Most significantly, these site-specific FP measurements provide insight into structural remodeling of αS fibrils by small molecules and have the potential for use in moderate-throughput screens to identify small molecules that could be used to treat Parkinson’s disease. © 2016 American Chemical Society.
- Published
- 2017