Fig. 1: PLA2G15 hydrolyses BMP lipids.
From: PLA2G15 is a BMP hydrolase and its targeting ameliorates lysosomal disease
a–c, Chemical structures of BMP (top) and phosphatidylglycerol (bottom) (a) and their degradation intermediates LPG (all possible structures) (b) and GPG (c). Red represents two glycerol groups, and blue represents the acyl chains. d, BMP hydrolase activity in the brain (left) and liver (right) lysates. The hydrolysis of 3,3′ 18:1-S,S BMP (1 µM) in buffers with pH range between 3.0 and 9.0 (Methods). Controls had similar reaction buffers, except with no lysate or BMP. Data are presented as mean ± s.d. of three biological replicates. e, Lysosomal lysates hydrolyse BMP with an acidic optimum. As in d but using brain lysosomal lysate (left) and liver lysosomal lysate (right). Data are presented as mean ± s.d. of n = 3. f, As in d but using brain and liver lysosomal lysates and in increasing concentration of amiodarone under acidic conditions (pH 5.0). Data are presented as mean ± s.d. of three biological replicates. g,h, PLA2G15 hydrolysed BMPs isolated from mouse liver lysosomes. g, Depiction of experimental design. h, Fold changes in the abundance of measured BMPs, and each time point was compared to the control. Data are presented as mean ± s.d. of five biological replicates. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001 by two-tailed unpaired t-tests for each BMP. i, Hydrolysis of 1 µM 3,3′ 18:1-S,S BMP by 100 nM recombinant wild-type PLA2G15 and S198A mutant under acidic conditions for 30-s reaction time. Data are presented as mean ± s.d. of three independent replicates. ****P = 0.000014 by two-tailed unpaired t-test. j, As in i using wild-type PLA2G15 in the presence of 20 µM amiodarone and fosinopril compared with the no inhibitor control. Data are presented as mean ± s.d. of three independent replicates. ****P < 0.0001 by two-tailed unpaired t-tests. a.u., arbitrary units; WT, wild type. Panel g was created using BioRender (https://biorender.com).
