TY - JOUR TI - Lysosomal cholesterol export reconstituted from fragments of Niemann-Pick C1 AU - Trinh, Michael Nguyen AU - Brown, Michael S AU - Seemann, Joachim AU - Goldstein, Joseph L AU - Lu, Feiran A2 - Prinz, William A A2 - Schekman, Randy VL - 7 PY - 2018 DA - 2018/07/26 SP - e38564 C1 - eLife 2018;7:e38564 DO - 10.7554/eLife.38564 UR - https://doi.org/10.7554/eLife.38564 AB - Niemann-Pick C1 (NPC1) is a polytopic membrane protein with 13 transmembrane helices that exports LDL-derived cholesterol from lysosomes by carrying it through the 80 Å glycocalyx and the 40 Å lipid bilayer. Transport begins when cholesterol binds to the N-terminal domain (NTD) of NPC1, which projects to the surface of the glycocalyx. Here, we reconstitute cholesterol transport by expressing the NTD as a fragment separate from the remaining portion of NPC1. When co-expressed, the two NPC1 fragments reconstitute cholesterol transport, indicating that the NTD has the flexibility to interact with the remaining parts of NPC1 even when not covalently linked. We also show that cholesterol can be transferred from the NTD of one full-length NPC1 to another NPC1 molecule that lacks the NTD. These data support the hypothesis that cholesterol is transported through interactions between two or more NPC1 molecules. KW - NPC1 fragments KW - cholesterol transport KW - reconstitution KW - split nanoluciferase KW - lysosome JF - eLife SN - 2050-084X PB - eLife Sciences Publications, Ltd ER -