title: Modulation of human α-synuclein aggregation by a combined effect of calcium and dopamine.
abstract: Parkinson's disease is characterized by the deposition of aggregated α-syn and its familial mutants into Lewy bodies leading to death of dopaminergic neurons. α-syn is involved in Ca(II) and dopamine (DA) signaling and their adequate balance inside neuronal cytoplasm is essential for maintaining healthy dopaminergic neurons. We have probed the binding energetics of Ca(II) and DA to human α-syn and its familial mutants A30P, A53T and E46K using isothermal titration calorimetry and have investigated the conformational and aggregation aspects using circular dichroism and fluorescence spectroscopy. While binding of Ca(II) to α-syn and its familial mutants was observed to be endothermic in nature, interaction of DA with α-syn was not detectable. Ca(II) enhanced fibrillation of α-syn and its familial mutants while DA promoted the formation of oligomers. However, Ca(II) and DA together critically favored the formation of protofibrils that are more cytotoxic than the mature fibrils. Using fluorescently labeled cysteine mutant A90C, we have shown that different aggregating species of α-syn formed in the presence of Ca(II) and DA are internalized into the human neuroblastoma cells with different rates and are responsible for the differential cytotoxicity depending on their nature. The findings put together suggest that an interplay between the concentrations of Ca(II), DA and α-syn can critically regulate the formation of various aggregating species responsible for the survival of dopaminergic neurons. Modulating this balance leading to either complete suppression of α-syn aggregation or promoting the formation of mature fibrils could be used as a strategy for the development of drugs to cure Parkinson's disease.
authors.affiliation: School of Biotechnology, Jawaharlal Nehru University, New Delhi 110 067, India.
authors.forename: Manish Kumar
authors.initials: MK
authors.lastname: Jain
authors.affiliation: School of Biotechnology, Jawaharlal Nehru University, New Delhi 110 067, India. Electronic address: rajivbhat@mail.jnu.ac.in.
authors.forename: Rajiv
authors.initials: R
authors.lastname: Bhat
mesh_terms: D002118:Calcium / Q000378:metabolism / Q000494:pharmacology*; D002151:Calorimetry; D045744:Cell Line, Tumor; D002942:Circular Dichroism; D004298:Dopamine / Q000378:metabolism / Q000494:pharmacology*; D006801:Humans; D018613:Microscopy, Confocal; D046529:Microscopy, Electron, Transmission; D009154:Mutation / Q000235:genetics; D009447:Neuroblastoma / Q000473:pathology; D011485:Protein Binding / Q000187:drug effects; D021381:Protein Transport / Q000235:genetics; D013816:Thermodynamics; D014162:Transfection; D014443:Tyrosine / Q000378:metabolism; D051844:alpha-Synuclein / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure
chemical_list: D051844:alpha-Synuclein; D014443:Tyrosine; D002118:Calcium; D004298:Dopamine
keywords: Amyloid; Calcium; Cytotoxicity; Dopamine; EGCG; Human α-syn