The PDZ domain of PDZGEF binds to phosphatidic acid.

<p>A. The PDZ domain of PDZGEF (CFP-PDZ) in HEK293T cells shows vesicular localization in the presence of PIP5Kα. B. Treatment of HEK293T cells with the PLD1 inhibitor CAY10593 impairs association of the PDZ domain (CFP-PDZ) with locally generated PA on PIP5Kα–generated vesicles. The bar graphs show the percentage of cells showing vesicular localization of the protein in the presence of PIP5Kα (8/11 cells for CFP-PDZ and 4/12 cells for CFP-PDZ in presence of CAY10593). All scale bars: 10 µm. C. Protein-lipid overlay assay of HA tagged PDZ domain of PDZGEF1 (PDZ) isolated from HEK293T cells using PIP strips containing 100pmol/spot of the following lipids: 1: lysophosphatidic acid (LPA), 2: lysophosphocholine (LPC), 3: phosphatidylinositol (PI), 4: phosphatidylinositol 3-phosphate [PI(3)P], 5: PI(4)P, 6: phosphatidylinositol 5-phosphate [PI(5)P], 7: phosphatidylethanolamine (PE), 8: phosphatidylcholine (PC), 9: sphingosine 1-phosphate (S1P), 10: phosphatidylinositol 3,4-bisphosphate [PI(3,4)P2], 11: phosphatidylinositol 3,5-bisphosphate [PI(3,5)P2], 12: phosphatidylinositol 1,2-bisphosphate [PI(4,5)P2], 13: phosphatidylinositol 3,4,5-trisphosphate [PI(3,4,5)P3], 14: phosphatidic acid (PA), 15: phosphatidylserine (PS) or 16: blue bank. Arrow indicates binding to PA.