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Only BlaP(Gln)55 and BlaP(Gln)79 form amyloid-like fibrils when incubated in the presence of 1.85 M urea.

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posted on 20.02.2013, 07:59 by Natacha Scarafone, Coralie Pain, Anthony Fratamico, Gilles Gaspard, Nursel Yilmaz, Patrice Filée, Moreno Galleni, André Matagne, Mireille Dumoulin

(A) Aggregation kinetics of 110 µM BlaP (blue), BlaP(Gln)23 (red), BlaP(Gln)30 (green), BlaP(Gln)55 (pink) and BlaP(Gln)79 (cyan) at 25°C in the presence of 1.85 M urea in PBS, pH 7.5, followed by measuring the concentration of protein remaining soluble. Time-points shown with an error bar are the average of three independent time-courses for BlaP(Gln)30, BlaP(Gln)55 and BlaP(Gln)79 and two independent time-courses for BlaP. Error bars show the standard deviations. Only one time-course was carried out with BlaP(Gln)23. (B) ThT fluorescence intensities at 482 nm in the presence of BlaP and chimeras samples at T0 (solid bars) and Tf (dashed bars). T0 and Tf correspond to the initial and final points of one time-course for each protein. Data are the average of three measurements and error bars represent the standard deviations. a.u., arbitrary units. (C) TEM images of the protein samples at Tf. The scale bar is 1 µm. (D) X-ray fibre diffraction patterns from BlaP(Gln)55 and BlaP(Gln)79 fibrils. White and black arrows indicate meridional and equatorial reflections at 4.7 Å and ca. 9.5 Å, respectively.