Ionic liquids promote amyloid formation from α-synuclein

Amyloid formation and disaggregation of α-synuclein and its tandem repeat (α-TR)

“…In a previous report, we have shown that ionic liquids could assist and stabilize the fibril formation process of a-synuclein [13]. Here, we examined the effects of two representative ionic liquids, 1-butyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide [BIMbF 3 Im] and 1-butyl-3-methylimidazolium tetrafluoroborate [BIMBF 4 ], on the fibril formation of a-synuclein and its tandem repeat (a-TR).…”

“…Single colonies of BL21 (DE3) were grown, and lyophilized and stored at À80°C until further use. The protein purities were judged to be >95% from SDS-PAGE and protein concentration was determined as described [13,15].…”

“…The time course fibrillizations of a-synuclein and its tandem repeat (a-TR) was measured by the ThT fluorescence assay [13]. Purified proteins were incubated at room temperature in 1.5-ml sterile polypropylene tubes with continuous shaking in the absence and presence of ionic liquids at a ratio (v/v) of 1%, 2%, or 5%.…”

“…10 lL aliquots of diluted samples were deposited on carbon-coated copper grids and stained with 2% (w/v) uranyl acetate solution for 1 min. Samples were air-dried and grids were examined in JEM1010 (JEOL, Japan) TEM at 40 kV, and digital images were acquired with a charge-coupled device camera [13].…”

“…To date, therapeutic strategies are mainly focused on the inhibition of the amyloid formation process, as early as its initial stage, by using organic molecules, heat shock proteins and antibodies [10][11][12]. Here, we suggest alternative strategies which avoid cytotoxic intermediates by using ionic liquids (ILs) that could stabilize the formation of amyloid fibrils [13,14]. As a first step, we have investigated the interaction of asynuclein with ionic liquids, using several biochemical techniques, including Thioflavin T assays and transmission electron microscopy (TEM).…”