RESUMO
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.
Assuntos
Catequina/análogos & derivados , Dopamina/química , Masoprocol/química , Agregados Proteicos/efeitos dos fármacos , Bibliotecas de Moléculas Pequenas/química , alfa-Sinucleína/química , Sequência de Aminoácidos , Catequina/química , Catequina/farmacologia , Dopamina/farmacologia , Polarização de Fluorescência , Corantes Fluorescentes/química , Humanos , Masoprocol/metabolismo , Fosfatidilcolinas/química , Proteínas Recombinantes/química , Bibliotecas de Moléculas Pequenas/farmacologia , Dodecilsulfato de Sódio/química , Lipossomas Unilamelares/química , Xantenos/químicaRESUMO
Controlled generation of singlet oxygen is very important due to its involvement in scheduled cellular maintenance processes and therapeutic potential. As a consequence, precise manipulation of singlet oxygen release rates under mild conditions, is crucial. In this work, a cross-linked polyacrylate, and a polydimethylsiloxane elastomer incorporating anthracene-endoperoxide modules with chain extensions at the 9,10-positions were synthesized. We now report that on mechanical agitation in cryogenic ball mill, fluorescence emission due to anthracene units in the PMA (polymethacrylate) polymer is enhanced, with a concomitant generation of singlet oxygen as proved by detection with a selective probe. The PDMS (polydimethylsiloxane) elastomer with the anthracene endoperoxide mechanophore, is also similarly sensitive to mechanical force.