Expression and Purification of Untagged α-Synuclein.

Expression and Purification of Untagged α-Synuclein.
Methods Mol Biol. 2019;1948:261-269
Authors: Powers AE, Patel DS
Abstract
α-Synuclein (αS) is an abundant neuronal protein which has been implicated, among others, in the pathogenesis of neurodegenerative diseases like Parkinson's disease (PD) and dementia with Lewy bodies (DLB). In fact, αS is the major constituent of Lewy bodies, the primarily proteinaceous inclusions found in the nervous tissue of PD and DLB patients. While its physiological role is unclear, it is believed to be involved in the regulation of synaptic vesicle exocytosis. However, in a disease state, αS will "misfold" and aggregate, leading to neuronal dysfunction and death. The study of the molecular events underlying pathogenesis, especially with biophysical and biochemical approaches, requires highly pure untagged αS. In this protocol we describe a method to purify untagged recombinant αS, which can be used for binding, folding, and aggregation studies. The purification method includes a cell lysis step, followed by two chromatography steps: ion-exchange chromatography first, and size-exclusion chromatography for polishing.
PMID: 30771184 [PubMed - indexed for MEDLINE]

Source: Mol Biol Cell - July 16, 2019 Category: Molecular Biology Authors: Powers AE, Patel DS Tags: Methods Mol Biol Source Type: research