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. 2024 May 2;12:RP90996. doi: 10.7554/eLife.90996

Table 2. The FRET (E) and Stoichiometry (S) of the FRET states of OpuA-K521C, their relative abundance and the transition rate constants between the states.

Table 2—source data 1. Burst variance analysis and mpH2MM of the FRET bursts of OpuA-K521C (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.
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
50 mM BIS-TRIS pH 7.0, 0 mM KCl 0.537 0.250 64.1 0.521 0.749 35.9 118 153 16,788
0 mM KCl 0.531 0.248 66.9 0.519 0.742 33.1 123 175 27,047
0 mM KCl, 100 µM glycine betaine 0.532 0.245 68.3 0.517 0.781 31.7 120 166 17,364
50 mM KCl 0.526 0.225 68.0 0.516 0.735 32.0 88 153 14,708
100 mM KCl 0.525 0.202 74.6 0.513 0.691 25.4 78 178 13,532
200 mM KCl 0.519 0.183 81.4 0.518 0.663 18.6 41 153 16,454
400 mM KCl 0.516 0.168 82.0 0.518 0.620 18.0 28 120 15,946
600 mM KCl 0.497 0.180 82.6 0.490 0.628 17.4 42 173 38,604
600 mM KCl (V149Q) 0.495 0.164 81.9 0.492 0.608 18.1 31 112 11,034
1000 mM KCl 0.505 0.148 79.9 0.511 0.591 20.1 26 109 12,666
600 mM KCl, 20 mM Mg-ATP 0.524 0.157 81.5 0.527 0.510 18.5 16 121 12,061
600 mM KCl, 20 mM Mg-ATP, 100 µM glycine betaine 0.515 0.155 78.5 0.511 0.507 21.5 0 52 17,155
600 mM KCl, 20 mM Mg-ATP, 100 µM glycine betaine (E190Q) 0.540 0.141 81.4 0.547 0.505 18.6 7 38 10,832
600 mM KCl, 20 mM Mg-ATP, 100 µM glycine betaine, 500 µM orthovanadate 0.517 0.154 78.9 0.513 0.529 21.5 17 87 21,052
600 mM KCl, 50 mM glutamate, 50 mM arginine 0.510 0.162 79.9 0.513 0.549 20.1 19 133 12,722
*

All conditions, except the first, contained 50 mM HEPES-K pH 7.0.