Melting temperature of MftR and mutant proteins
determined by differential
scanning fluorometry. Thermal denaturation is represented by the fluorescence
intensity resulting from the binding of SYPRO Orange to denatured
protein as a function of temperature. (A) Thermal denaturation profile
of WT MftR. (B) Thermal denaturation profile of D56S, R63S, and R89N.
(C) Melting profile of W11F; a higher initial fluorescence may reflect
the presence of already denatured protein. (D) Melting profile of
WT with the addition of 100 μM urate. (E) Denaturation profile
of D56S, R63S, and R89N with urate. (F) Denaturation profile of W11F
with urate. (G) Thermal denaturation profile of MftR with 6 μM mftpO. (H) Denaturation profile of mutant D56S, R63S, and
R89N with mftpO. (I) Symbols used in panels A–H
for MftR and mutants.