Table 3. The FRET (E) and Stoichiometry (S) of the FRET states of OpuA-N414C, their relative abundance, and the transition rate constants between the states.
Table 3—source data 1. Burst variance analysis and mpH2MM of the FRET bursts of OpuA-N414C (variants) in different buffers.
(A) From top to bottom: (1) Burst variance analysis of the bursts which were corrected by the leakage, crosstalk, and γ-correction factors and which were selected after removing donor-only and acceptor-only bursts. The standard deviation of FRET in each burst is plotted against its mean FRET. Black squares show average values per FRET bin. Black dotted line shows the expected standard deviation in the absence of within-burst dynamics. (2) 2D E-S histogram shows the same data as in (1), with on both sides a histogram that represents the same bursts. (B) Plot of the ICL-values for each final model. The model used in the downstream analysis and following figures is shown as a star. (C) Plot of the BIC’-values for each final model. The red line represents a 0.05 cut-off. The model used is shown as a star. (D) Burst-based 2D E-S scatter plot. Bursts are colored on the basis of the assigned state of the chosen mpH2MM model. If a burst contains more than one state, it is assigned as being dynamic. (E) Dwell-based 2D E-S scatter plot. Dwells are colored on the basis of the assigned state of the chosen mpH2MM model. The dwells were corrected for leakage, direct excitation and the γ-factor. Black dots in A, D and E represent the average value of each state.
elife-90996-table3-data1.zip (10.4MB, zip)
| Condition* | Low-FRET state (L) | High-FRET state (H) | Transition rate (s–1) | Number of FRET dwells | |||||
|---|---|---|---|---|---|---|---|---|---|
| S | E | % dwells | S | E | % dwells | L-H | H-L | ||
| 0 mM KCl | 0.527 | 0.099 | 72.2 | 0.543 | 0.509 | 27.8 | 15 | 68 | 12,162 |
| 600 mM KCl | 0.483 | 0.100 | 88.5 | 0.476 | 0.497 | 11.5 | 24 | 193 | 18,130 |
| 600 mM KCl, 20 mM Mg-ATP | 0.548 | 0.090 | 94.4 | 0.536 | 0.456 | 5.6 | 2 | 141 | 10,447 |
| 600 mM KCl, 20 mM Mg-ATP, 100 µM glycine betaine | 0.538 | 0.087 | 95.7 | 0.555 | 0.499 | 4.7 | 2 | 59 | 14,898 |
| 600 mM KCl, 20 mM Mg-ATP, 100 µM glycine betaine (E190Q) | 0.479 | 0.106 | 94.8 | 0.464 | 0.540 | 5.2 | 25 | 317 | 8024 |
| 600 mM KCl, 20 mM Mg-ATP, 100 µM glycine betaine, 500 µM orthovanadate | 0.538 | 0.085 | 94.6 | 0.578 | 0.568 | 5.4 | 5 | 186 | 7236 |
*
All conditions contained 50 mM HEPES-K pH 7.0.