Figure 2. DEER analysis of spin-labeled TM287/288.
Q-band background-corrected DEER traces [F(t)/F(0)] with fitted distribution function (left) and corresponding distance distribution (right) calculated using DeerAnalysis2015 and normalized by area. Traces are shown for the six spin-labeled pairs in the absence of nucleotides (apo, cyan), ATP-EDTA (green), ATP-Mg incubated for 20 s (red) and ATP-Vi-Mg (magenta). (A and B) Extracellular pairs. (C and D) Intracellular pairs. (E) NBD pair 460TM287/363TM288 monitoring the consensus site. (F) NBD pair 350TM287/475TM288 monitoring the degenerate site. Transparent cyan and magenta rectangles outline the range of experimental distances characteristic of the IF and OF conformations, respectively. All primary DEER data can be found in Figure 2—figure supplement 1. The DEER data of the apo states presented in panels A, C, E, F are taken from (Hohl et al., 2014). Traces detected after incubation with 14 mM ATP and 2.5 mM EDTA are shown in Figure 2—figure supplement 4.
Figure 2—figure supplement 1. Primary DEER traces of spin-labeled pairs in wildtype TM287/288.
Figure 2—figure supplement 2. DEER analysis of TM287/288 in proteoliposomes.
Figure 2—figure supplement 3. DEER analysis of spin-labeled pairs in wildtype TM287/288 with additional nucleotide analogs and ADP-Mg.
Figure 2—figure supplement 4. DEER analysis of wildtype TM287/288 in the presence of 2.5 mM or 14 mM ATP and 2.5 mM EDTA.
