Aberrant maturation of mutant perforin underlies the clinical diversity of hemophagocytic lymphohistiocytosis
J. Clin. Invest. Kimberly A. Risma, et al. 116:182 doi:10.1172/JCI26217 [Go to this article.]

Figure 7
Western blot analysis of RBL-1 expressing PRF1 missense mutations with reduced processing to the mature form. The maturation of perforin was interrupted by alkalinization of acidic compartments with CMA or by inhibition of proteolysis by leupeptin overnight. Under nonreducing SDS-PAGE conditions, immunoblotting (H315 antibody) revealed that mutant proteins (PRF1-A91V, -R232H, -R361W, and -G429E) appeared to have a maturation pattern comparable to that of WT perforin, despite reduced intensity of the mature band: CMA completely blocked processing to the mature band, while leupeptin blocked it only partially. Twenty micrograms protein was loaded per lane, except for the WT perforin, of which 10 μg was loaded because of the strength of the signal.