Fig. 7
From: The revealing of a novel lipid transfer protein lineage in green algae
Protein-lipid overlay (PLO) assay and proteinase digestion analysis of CrLTP2. A PLO assay. The binding ability of CrLTP2 protein to the pre-spotted lipid materials were tested. A membrane dotted with the lipid ligands and a duplicated control membrane are respectively reacted with the purified CrLTP2 protein (CrLTP2) and without the protein (control) and immunodetected with an anti-CrLTP2 antibody. The dotted circles depict the position of the pre-spotted lipid materials, as indicated: 1, Phenylalanine; 2, cinnamic acid; 3, p-coumaric acid; 4, caffeic acid; 5, ferulic acid; 6, capric acid (C10:0); 7, lauric acid (C12:0); 8, myristic acid (C14:0); 9, palmitic acid (C16:0); 10, stearic acid (C18:0); 11, arachidic acid (C20:0); 12, behenic acid (C22:0); 13, lignoceric acid (C24:0); 14, 12-hydroxystearic acid; 15, 15-hydroxypentadecanoic acid;16, 16-hydroxyhexadecanoic acid. B The analysis of the protease-resistant ability of CrLTP2 protein. Bovine serum albumin (BSA), the control substrates, and CrLTP2 protein were mixed, incubated with protease and collected at different time points: 0, 5, 10, 15, 30, 45, and 60 minutes. The digested protein products were resolved using SDS-PAGE and stained with Coomassie Brilliant Blue dye. Cropped blot images are shown, and full length blots are presented in Supplementary Fig. 7. The numbers on the left indicate the protein molecular masses in kDa, and the arrows on the right indicate the positions of BSA and CrLTP2