Skip to main content
. Author manuscript; available in PMC: 2021 Oct 15.
Published in final edited form as: Cell Chem Biol. 2020 Aug 11;27(10):1296–1307.e5. doi: 10.1016/j.chembiol.2020.07.015

Figure 2.

Figure 2.

The presence of BMP and its abundance are primary determinants of leakage outcome. A) Effect of lipid composition on 3TAT-mediated leakage. LUVs were prepared with a lipid composition of X:PC:PE (77:19:4), X being either the zwitterionic lipid PC or the 3 anionic regioisomers PG, 3,3’-BMP and 2,2’-BMP. The fatty acid present in all lipids was oleic acid [18:1]. The release of free calcein upon treatment with 3TAT at the peptide:lipid ratios indicated was measured for each LUV suspensions. The fraction of LUV-bound peptide is 100% for all LUVs at 1:50 peptide:lipid ratios (as determined by sedimentation). B) Effect of the abundance of 3,3’-BMP in the bilayer on membrane leakage. LUVs were prepared with a lipid composition of BMP:PC:PE (X:96-X:4), with X being varied from 10% to 77%. LUVs exclusively containing 3,3’-BMP were also prepared. Free calcein leakage was quantified at the 3TAT:lipid ratios indicated. C) Effect of pH, cholesterol and buffer composition on leakage. The calcein leakage of 3,3’-BMP LUVs (BMP:PC:PE 77:19:4 or BMP:Chol:PE 77:19:4) treated with 3TAT was determined in PBS buffer at a pH of either 5.5 or 7.4, or in Tris buffer at pH 5.5.