Identification of pharmacological chaperones as potential therapeutic agents to treat phenylketonuria
J. Clin. Invest. Angel L. Pey, et al. 118:2858 doi:10.1172/JCI34355 [Go to this article.]

Figure 4
Effect of hit compounds III and IV on the stability and function of tetrameric PAH proteins. (A) SEC determination of the fraction of remaining tetramer in WT and mutant proteins after heat shock at 50°C for 20 minutes in the absence or presence of 100 μM of compound III and IV in 2.5% DMSO (also present in control sample). Data are expressed as area of the tetrameric fraction after heat shock related to the area in corresponding samples incubated for 10 minutes at 25°C. Inset shows representative SEC profiles obtained for tetrameric WT-PAH incubated at 25°C for 10 minutes (solid line) or for 2 independent samples treated at 50°C for 20 minutes with (dashed line) or without (dotted lines) filtering through a 0.22-μm pore filter to remove protein aggregates. Only the aggregated forms are eliminated by filtration. Elution volumes are approximately 15 ml for tetrameric forms and approximately 10 ml for aggregates (the void volume of the column). (B–D) Thermal inactivation experiments (B, WT-PAH; C, I65T-PAH; D, R68S-PAH) performed by 10-minute incubation at the indicated temperatures without (circles) or with 100 μM of compound III (triangles) and IV (squares) and subsequent measurement of PAH activity at 25°C. Data are expressed as fractional activity after incubation at different temperatures related to the corresponding activity after incubation at 25°C and are given as mean ± SD of triplicate experiments.