Fig. 2
From: A thiol probe for measuring unfolded protein load and proteostasis in cells
TPE-MI preferentially reacts with buried cysteine thiols in unfolded proteins and switches on fluorescence. Shown are representative reaction kinetics for four proteins (note that the absolute fluorescence values between graphs here and in other figures cannot be compared to each other due to differences in instrument settings). a Bovine β-lactoglobulin, which contains five thiols: one buried free thiol and four disulphide-linked thiols; and bovine ubiquitin, which contains no thiols. b Saccharomyces cerevisiae enolase, which contains one buried free thiol and c human peroxiredoxin 3, which contains three thiols: one buried free thiol and two disulphide-linked surface-exposed thiols. Proteins were suspended in 100 mM sodium phosphate, pH 7.4, alone or supplemented with guandine hydrochloride (GuHCl) to induce denaturation. At the start of the reaction 50 μM TPE-MI was added. d Same design but with 50 μM N-methylmaleimide (NMM) added before the addition of TPE-MI.
