Q-band background-corrected DEER traces [F(t)/F(0)] with fitted distribution function (left) and corresponding distance distribution (right) for the six spin-labeled pairs engineered into wildtype TM287/288. The upper distance distributions correspond to those presented in
Figure 2, the bottom distributions to the additional nucleotide analogs and ADP-Mg. (
A and
B) Extracellular pairs 150
TM287/295
TM288 and 50
TM287/271
TM287. (
C and
D) Intracellular pairs 131
TM288/248
TM288 and 231
TM287/304
TM287. (
E) NBD pair 460
TM287/363
TM288 monitoring the consensus site. (
F) NBD pair 350
TM287/475
TM288 monitoring the degenerate site. The data of the apo and AMP-PNP-Mg states of the four pairs 150
TM287/295
TM288, 131
TM288/248
TM288, 460
TM287/363
TM288, 350
TM287/475
TM288 in the wildtype transporter are taken from (
Hohl et al., 2014).