Effect of ATP on the surface hydrophobicity of M. leprae HSP18.
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First, HSP18 was incubated in absence (solution 1) and presence of presence of 0.01 mM ATP (solution 6)/1 mM ATP (solution 2)/1 mM ATP-γS (solution 5) at 25°C for 1 hr. Then, part of solution 1 and 2 were dialysed overnight against 50 mM phosphate buffer, pH 7.5. The resultant solution achieved from the dialysis of solution 1 and 2 were termed as solution 3 and 4, respectively. All these solutions were incubated with 10 μM bis-ANS at 25°C for 1 hr. The protein concentration was 0.05 mg/ml. The fluorescence spectrum of bis-ANS bound to different samples was recorded in the range 450–550 nm at 25°C. The excitation wavelength was 390 nm. Trace 1: bis-ANS + solution 1; Trace 2: bis-ANS + solution 6; Trace 3: bis-ANS + solution 2; Trace 4: bis-ANS + solution 5; Trace 5: bis-ANS + solution 3; Trace 6: bis-ANS + solution 4.