Journal: PLoS ONE

Article Title: FYVE-Dependent Endosomal Targeting of an Arrestin-Related Protein in Amoeba

doi: 10.1371/journal.pone.0015249

Figure Lengend Snippet: ( A ) Lipid overlay assay. AdcA FYVE domain was expressed as a MBP-fusion protein and tested for its binding to lipids immobilized on a membrane. MBP alone was used as a control. LPA, lysophosphatidic acid; LPC, lysophosphocholine; PI, phosphatidylinositol; PI3P, phosphatidylinositol 3-phosphate; PI4P, phosphatidylinositol 4-phosphate; PI5P, phosphatidylinositol 5-phosphate; PE, phosphatidylethanolamine; PC, phosphatidylcholine; S1P, sphingosine 1-phosphate; PI3,4P 2 , phosphatidylinositol 3,4-bisphosphate; PI3,5P 2 , phosphatidylinositol 3,5-bisphosphate; PI4,5P 2 , phosphatidylinositol 4,5-bisphosphate; PI3,4,5P 3 , phosphatidylinositol 3,4,5-trisphosphate; PA, phosphatidic acid; PS, phosphatidylserine; no, blank. ( B ) Binding of Ins(1,3)P 2 . Variations of tryptophan intrinsic fluorescence (λ ex 290 nm) of MBP-FYVE (o) and MBP (•) were measured between 300 and 400 nm in response to addition of Ins(1,3)P 2 , a soluble analog of PI(3)P. Variations were expressed as ΔF/F 0 in percent. ( C ) AdcA membrane association is pH dependent. Cells expressing AdcA GFP were broken in different buffers at different pHs and treated as in panel ( A ).

Article Snippet: PIP strips and Ins(1,3)P 2 were purchased from Echelon Biosciences Inc. (Tebu-bio, Le-Perray-en-Yvelines, France).

Techniques: Overlay Assay, Binding Assay, Fluorescence, Expressing