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. Author manuscript; available in PMC: 2014 Jan 1.
Published in final edited form as: Biochim Biophys Acta. 2012 Jul 20;1834(1):464–469. doi: 10.1016/j.bbapap.2012.06.018

Figure 6.

Figure 6

Hydroxyl radical modified PTP1B can be reactivated by thiols. PTP1B with final concentration of 2 μM was incubated in 100 μl 50 mM 3,3-dimethylglutarate buffer (pH 6.0) containing 50 μM H2O2. When it was completely inactivated, 5 μl of the inactivated PTP1B solution was added to the reactivation system in a 1 ml-cuvette which was pre-incubated at 25°C; and the final concentration of PTP1B was 10 nM. The reactivation system was 50 mM 3,3-dimethylglutarate buffer (pH 6.0), 20 mM pNPP, and 10 mM DTT (for the upper curve), or 20 mM GSH (for the lower curve). The reaction was measured by following the change of optical density at 405 nm upon addition of the enzyme at 25°C.