October 17, 2014
Optimal Recruitment of Adaptor Protein 1 to the Surface of Giant Unilamellar Vesicles
Clathrin-mediated endocytosis (CME) is the process by which cells uptake molecules from the extracellular environment through membrane invagination. CME requires the help from many different proteins, and is crucial for healthy cells to communicate, function properly, and survive. Therefore, abnormalities in CME can result in harmful effects, such as virus invasion, muscular dystrophy, and neuropathies such as Huntington disease. Adaptor proteins (AP) are essential for the proper selection of extracellular cargo, and specifically AP1 is necessary for the selection of materials for transport between the trans-Gogli network (TGN) and the endosomes. This summer, I investigated the optimal conditions required for maximal recruitment of AP1 to the membrane of giant unilamellar vesicles (GUVs). GUVs were produced to mimic the membrane of the TGN, and the composition of the lipid membrane was varied to test different conditions. AP1 recruitment was quantified by SDS-PAGE and western blot techniques, and further analyzed using Image J and OriginPro software.
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