Extended Data Fig. 3: Purification and characterization of lysosomal phospholipases.
From: PLA2G15 is a BMP hydrolase and its targeting ameliorates lysosomal disease
a, Purification of recombinant His-tagged PLA2G15 S198A. Representative image of 47.5 kDa PLA2G15 S198A-6xHis protein purified to homogeneity and stained with Coomassie Blue. b, Immunoblot analysis of PLA2G15 S198A using anti-His antibody. c, Melting temperature (Tm) curves indicate that wildtype and S198A mutant maintain thermal stability. Tm is indicated on the graph. d, Purification of recombinant His-tagged PLBD2. Representative image of PLBD2-6xHis protein purified to homogeneity and stained with Coomassie Blue. The three bands are indicative of 66.5 kDa proenzyme, 46.5 kDa matured form and 33 kDa N-terminus pro-domain. e, Immunoblot analysis of PLBD2 using anti-His antibody. f, PLBD2 has no BMP hydrolase activity. BMP Hydrolase activity by 100 nM wildtype recombinant PLA2G15 and PLBD2 proteins under acidic conditions determined using LC-MS. Data are mean ± s.d. of n = 3 independent replicates from each protein. ****p = 0.0000021 using two-tailed unpaired t-tests. Gel source data are provided in Supplementary Fig. 1.
