TIP30 and Endo B1 strongly bind phosphatidic acid.

(A) Fatty acylation of endosomal lipids by the TIP30 complex. [3H]-arachidonic acid can be transferred to endosomal membrane lipids by the TIP30 complex, but not by TIP30M immunoprecipitates or control immunoprecipitates (left panel). The transfer was blocked by 10 µM triacsin C (right panel). Image was acquired by scanning lipids resolved on TLC plate with a Molecular Dynamics Storm 860. * indicates the radiolabeled lipid. (B) The schematic diagram shows the lipid species pre-spotted on membranes that are used in lipid-protein overlay assays. (C) TIP30 and Endo B1 strongly bind phosphatidic acid (PA). Protein-lipid overlay assays were carried out by incubating recombinant proteins with membrane strips containing 15 pre-spotted lipids. Membranes were scanned using a Li-Cor scanner after being sequentially overlaid with primary and fluorescent secondary antibodies.