Fig. 2.

Effects of GSH on the folding of Der p 1. (A) SDS-PAGE analysis (stained with Coomassie blue) of the effect of GSH (1 mM) on the relative migration distance of Der p 1, with and without GSTpi. Untreated- and DTT-treated proteins were used as controls. Arrows indicate the positions of Der p 1 and the GSTpi. C, untreated Der p 1 used as control; M, molecular masses marker. (B) Redox fluorescence switch (RFS) assay of Der p 1. Der p 1 was incubated with 1 mM GSH for 30 min and then successively with 50 mM NEM, 2.5 mM DTT or fluorescent maleimide (maleimide with BODIPY-FL) as indicated. BODIPY-FL labelling after NEM addition without DTT measures non-specific labelling (negative control). The amount of loaded-protein was analysed with SYPRO staining. A representative example of three independent experiments is shown.