Spectroscopic and calorimetric studies of congo red dye-amyloid peptide complexes


Thermodynamic properties of complexes of Con ?go Red (CR) dye with amyloid ? (A?) peptides were studied by both absorption spectroscopy and isothermal titration calorimetry (ITC). Based on the absorption spectrum for the formation of CRAβ complexes in phosphate buffered saline solution (pH 7.4), van’t Hoff plots over a temperature range of 10oC to 70oC were created for CRAβ140, Aβ1228, and Aβ142. The plot for CR Aβ1228 complex showed a relatively linear feature within the given temperature range with ?H = –10.1 ?0.6 kJ/mol and ?S = + 0.128 ? 0.002 kJ/(mol K). However, the plot for CRAβ140 and CRAβ142 complexes exhibited two distinct linear regions with opposite slopes centered at a specific temperature, Ts, which was 54.7 ? 0.2℃ and 34.8 ? 0.2℃, respectively. The ITC experiments conducted at 25℃in water exhibited quite a different situation from the above mentioned spectroscopic approach. The ITC studies yielded a ?H of –85.3 ? 0.2 kJ/mol for the CRAβ1228 complex with negative entropy change –0.152 kJ/mol K). For CRAβ140, the ITC studies indicated the presence of two binding sites with ?H1 = –81.8 ? 0.3 kJ/mol and ?H2 = –119.5 ? 0.2 kJ/mol with K1 = 5.5 ? 0.7 ? 106 M1 and K2 = 6.9 ? 2.4 ? 108 M1, respectively. These binding constants are consistent with the model suggested by several studies. Both binding sites showed negative entropy changes suggesting that the formation of the complex is enthalpically driven. The disagreement in thermochemical values between two different methods confirmed that the enthalpy and entropy are heavily dependent on temperature and buffer/salt environment, and may involve inherently different reaction paths.

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Yokoyama, K. , Fisher, A. , Amori, A. , Welchons, D. and McKnight, R. (2010) Spectroscopic and calorimetric studies of congo red dye-amyloid peptide complexes. Journal of Biophysical Chemistry, 1, 153-163. doi: 10.4236/jbpc.2010.13018.

Conflicts of Interest

The authors declare no conflicts of interest.


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