Table 1.
Fluorescence | Circular dichroism | |||||||
Total dimer concentration | ΔG0.1a | m1b | ΔG0.2a | m2b | ΔG0.1a | m1b | ΔG0.2a | m2b |
Dimeric intermediate | ||||||||
10−7 M | 6.06 ± 5.70 | 1.90 ± 1.83 | 13.55 ± 8.36 | 1.20 ± 1.44 | ndc | nd | nd | nd |
2.10−7 M | 5.97 ± 14.97 | 1.74 ± 3.52 | 10.76 ± 14.37 | 0.51 ± 1.71 | nd | nd | nd | nd |
10−6 M | 8.09 ± 1.93 | 2.34 ± 0.55 | 14.01 ± 4.01 | 1.42 ± 0.72 | 9.16 ± 2.71 | 2.66 ± 0.77 | 12.93 ± 3.05 | 1.23 ± 0.55 |
4.10−6 M | 8.9 ± 2.4 | 2.3 ± 0.6 | 14.0 ± 6.9 | 1.4 ± 1.2 | 12.19 ± 9.34 | 3.12 ± 2.22 | 10.61 ± 6.10 | 0.91 ± 0.98 |
Monomeric intermediate | ||||||||
10−7 M | 16.23 ± 1.14 | 2.31 ± 0.39 | 5.51 ± 0.62 | 1.26 ± 0.06 | nd | nd | nd | nd |
2.10−7 M | 18.16 ± 0.28 | 2.94 ± 0.05 | 6.43 ± 0.50 | 1.26 ± 0.12 | nd | nd | nd | nd |
10−6 M | 18.55 ± 0.11 | 3.14 ± 0.04 | 6.21 ± 0.16 | 1.29 ± 0.02 | 18.47 ± 0.13 | 3.12 ± 0.02 | 6.12 ± 0.04 | 1.28 ± 0.02 |
4.10−6 M | 18.26 ± 0.66 | 3.00 ± 0.18 | 6.10 ± 0.49 | 1.21 ± 0.05 | 18.72 ± 0.74 | 3.14 ± 0.22 | 6.14 ± 1.17 | 1.30 ± 0.15 |
The unfolding transitions at different protein concentrations were individually fitted to both equation 2 (dimeric intermediate) and equation 3 (monomeric intermediate) as developed in Materials and Methods. The errors correspond to standard deviations obtained from the fits.
a In kCal • mole−1.
b In kCal • mole−1 • M−1.
c nd, not determined.