Table 1.
Comparison of parameters of distributions for three intramolecular distances in the Escherichia coli AK molecule
| AK mutanta |
PF(r)b |
PC(r) double-kineticsc |
PC(r) intensityd |
kF (s−1)g | |||
|---|---|---|---|---|---|---|---|
| Peak (Å)e | FWHM (Å)f | Peak (Å) | FWHM (Å) | Peak (Å) | FWHM (Å) | ||
| 79(Sal)-86(Trp) | 20.7 (20.0–20.9) | 6 (4–8) | 16.6 (16.1–20.5) | 19 (5–20) | 17.8 (17.7–18.7) | 20 (12–21) | 0.40 (0.31–0.47) |
| 79(Cou)-86(Trp) | 20.7 (20.0–20.9) | 6 (4–8) | 15.2 (15.2–21.7) | 18 (6–18) | 14.7 (14.4–16.4) | 17 (13–18) | 0.56 (0.50–0.62) |
| 28(Sal)-71(Trp) | 17.2 (16.4–17.7) | 10 (8–12) | 20.6 (17.7–21.8) | 12(9–17) | 19.2 (17.6–20.1) | 13 (5–21) | 0.63 (0.60–0.66) |
| 18(Sal)-203(Trp) | 15.2 (14.2–15.9) | 9 (8–11) | 34.9 (32.2–38.1) | 23 (16–31) | 23.3 (20.2–26.1) | 27 (9–30) | 0.43 (0.42–0.45) |
The comparisons were performed at the initial phase of refolding determined by analysis of the emission intensity detected kinetics, and by double-kinetics methods. AK, adenylate kinase; FWHM, full width at half-maximum.
The two labeled residues and the probes used as FRET donor and acceptor.
The final (folded) state intramolecular distribution obtained by trFRET measurements at 0.3 M GndHCl.
The initial (collapsed) state intramolecular distance distribution obtained by double-kinetics FRET measurements at the first few milliseconds of folding.
The initial (collapsed) state distribution obtained as extrapolation to zero-time from best-fit results of the combined analysis.
Most probable distance out of a skewed Gaussian radial distance distribution model.
The FWHM of a skewed Gaussian radial distance distribution model.
The first-order apparent refolding rate constant.