Kinetics of GSTP1-1 nitrosation monitored by intrinsic fluorescence.
A, representative transients showing the decrease in fluorescence when 1 μm of different cysteine mutants of GSTP1-1 was mixed with 500 μm nitrosating agent (GSNO or CysNO) at 37 °C. Curves were normalized to have the same starting fluorescence. B, S-nitrosation of 1 μm wild-type GSTP1-1 with 500 μm GSNO. The data were fit to a triple-exponential function. The residuals show an excellent fit to the data when the three exponential terms are fixed to the observed rate constants for nitrosation at Cys47, Cys101, and the alternate site, respectively, determined from fitting the data in B.