title: Sequential Extraction of Soluble and Insoluble Alpha-Synuclein from Parkinsonian Brains.
abstract: Alpha-synuclein (α-syn) protein is abundantly expressed mainly within neurons, and exists in a number of different forms - monomers, tetramers, oligomers and fibrils. During disease, α-syn undergoes conformational changes to form oligomers and high molecular weight aggregates that tend to make the protein more insoluble. Abnormally aggregated α-syn is a neuropathological feature of Parkinson's disease (PD), dementia with Lewy bodies (DLB) and multiple system atrophy (MSA). Biochemical characterization and analysis of insoluble α-syn using buffers with increasing detergent strength and high-speed ultracentrifugation provides a powerful tool to determine the development of α-syn pathology associated with disease progression. This protocol describes the isolation of increasingly insoluble/aggregated α-syn from post-mortem human brain tissue. This methodology can be adapted with modifications to studies of normal and abnormal α-syn biology in transgenic animal models harbouring different α-syn mutations as well as in other neurodegenerative diseases that feature aberrant fibrillar deposits of proteins related to their respective pathologies.
authors.affiliation: Reta Lila Weston Institute of Neurological Studies, UCL Institute of Neurology; rina.bandopadhyay@ucl.ac.uk.
authors.forename: Rina
authors.initials: R
authors.lastname: Bandopadhyay
mesh_terms: D001921:Brain / Q000378:metabolism* / Q000473:pathology; D001923:Brain Chemistry; D006801:Humans; D009474:Neurons / Q000378:metabolism / Q000473:pathology; D010300:Parkinson Disease / Q000378:metabolism* / Q000473:pathology; D066263:Protein Aggregation, Pathological; D014461:Ultracentrifugation / Q000379:methods; D051844:alpha-Synuclein / Q000302:isolation & purification*
chemical_list: D051844:alpha-Synuclein