Author: Rathore, Shailendra S.; Liu, Yinghui; Yu, Haijia; Wan, Chun; Lee, MyeongSeon; Yin, Qian; Stowell, Michael H.B.; Shen, Jingshi
Title: Intracellular Vesicle Fusion Requires a Membrane-Destabilizing Peptide Located at the Juxtamembrane Region of the v-SNARE Document date: 2019_12_24
ID: pudp1eoo_36
Snippet: Liposome leakage assays-Sulforhodamine B-loaded protein-free liposomes were mixed with buffer or a SNARE peptide (added to a final concentration of 100 μM). Sulforhodamine B fluorescence was measured over time at 37°C. At the end of the reactions, 10 μL of 10% CHAPSO was added to lyse the liposomes to obtain the maximum fluorescence. The data are shown as percentage of maximum fluorescence. The N-peptide and Vc peptide were expressed and purif.....
Document: Liposome leakage assays-Sulforhodamine B-loaded protein-free liposomes were mixed with buffer or a SNARE peptide (added to a final concentration of 100 μM). Sulforhodamine B fluorescence was measured over time at 37°C. At the end of the reactions, 10 μL of 10% CHAPSO was added to lyse the liposomes to obtain the maximum fluorescence. The data are shown as percentage of maximum fluorescence. The N-peptide and Vc peptide were expressed and purified from E. coli whereas the juxtamembrane motif peptide of VAMP2 was synthesized by Biometik (95% purity). The sequences of the SNARE peptides are listed below: N-peptide (residues 1-35 of syntaxin-1): MKDRTQELRTAKDSDDDDDVTVTVDRDRFMDEFFE.
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