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Detecting phospholipase activity with the amphipathic lipid packing sensor motif of ArfGAP1

Yunes et al Quartino. BBRC. 2018

The amphipathic lipid packing sensor (ALPS) motif of ArfGAP1 brings this GTPase activating protein to membranes of high curvature. Phospholipases are phospholipid-hydrolyzing enzymes that generate different lipid products that alter the lateral organization of membranes. Here, we evaluate by fluorescence microscopy how in-situ changes of membrane lipid composition driven by the activity of different phospholipases promotes the binding of ALPS. We show that the activity of phospholipase A2, phospholipase C and phospholipase D drastically enhances the binding of ALPS to the weakly-curved membrane of giant liposomes. Our results suggest that the enzymatic activity of phospholipases can modulate the ArfGAP1-mediated intracellular traffic and that amphiphilic peptides such as the ALPS motif can be used to study lipolytic activities at lipid membranes.

Authors: Quartino PY, Fidelio GD, JB Manneville, Goud B, Ambroggio EE.

Article: Detecting phospholipase activity with the amphipathic lipid packing sensor motif of ArfGAP1. Quartino PY, Fidelio GD, JB Manneville, Goud B, Ambroggio EE. Biochem Biophys Res Commun. 2018 Oct 20;505(1):290-294. doi: 10.1016/j.bbrc.2018.09.116. Epub 2018 Sep 22.