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. 2021 Jan 6;120(3):476–488. doi: 10.1016/j.bpj.2020.12.019

Figure 4.

Figure 4

Characterization of the IAPP-DnaK interactions by ensemble and single-molecule techniques. (A) Ensemble fluorescence anisotropy and (B) hydrodynamic radii (Rh) from FCS measurements of 400 nM TMR-IAPP in presence of 0–5 μM DnaK are shown. The symbols represent data. The solid line in (A) is guide to the eye. (C) Rh of 110 nM TMR-IAPP in presence of 1 μM of DnaK or Hsc70 under various solution conditions is shown. (D) Presence of oligomeric TMR-IAPP is reflected in the fluorescence bursts in the photon count traces obtained from four identical FCS measurements on the 400 nM TMR-IAPP sample is shown. The solutions are prepared in 10 mM phosphate buffer (pH 7.4) containing 150 mM NaCl, 1 mM EDTA, and 2 mM azide and incubated for O/N at RT. In some of the samples, as mentioned in (C), NaCl and EDTA have been substituted by 100 mM KCl and 5 mM MgCl2. The experiments have been performed four times. To see this figure in color, go online.