title: Role of Niemann-Pick type C1 protein in intracellular trafficking of low density lipoprotein-derived cholesterol.
abstract: Niemann-Pick type C (NPC) is a disease that affects intracellular cholesterol-trafficking pathways. By cloning the hamster ortholog of NPC1, we identified the molecular lesions in two independently isolated Chinese hamster ovary cell mutants, CT60 and CT43. Both mutants lead to premature translational terminations of the NPC1 protein. Transfecting hamster NPC1 cDNA complemented the defects of the mutants. Investigation of the CT mutants, their parental cells, and an NPC1-stable transfectant allow us to present evidence that NPC1 is involved in a post-plasma membrane cholesterol-trafficking pathway. We found that the initial movement of low density lipoprotein (LDL)-derived cholesterol to the plasma membrane (PM) did not require NPC1. After reaching the PM and subsequent internalization, however, cholesterol trafficking back to the PM did involve NPC1. Both LDL-derived cholesterol and cholesterol originating from the PM accumulated in a dense, intracellular compartment in the CT mutants. Cholesterol movement from this compartment to the PM or endoplasmic reticulum was defective in the CT mutants. Our results functionally distinguish the dense, intracellular compartment from the early endocytic hydrolytic organelle and imply that NPC1 is involved in sorting cholesterol from the intracellular compartment back to the PM or to the endoplasmic reticulum.
authors.affiliation: Department of Biochemistry, Dartmouth Medical School, Hanover, New Hampshire 03755, USA.
authors.forename: J C
authors.initials: JC
authors.lastname: Cruz
authors.forename: S
authors.initials: S
authors.lastname: Sugii
authors.forename: C
authors.initials: C
authors.lastname: Yu
authors.forename: T Y
authors.initials: TY
authors.lastname: Chang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D016466:CHO Cells; D002352:Carrier Proteins; D002462:Cell Membrane / Q000378:metabolism; D002499:Centrifugation, Density Gradient; D008078:Cholesterol, LDL / Q000378:metabolism*; D003001:Cloning, Molecular; D006224:Cricetinae; D004705:Endocytosis; D004721:Endoplasmic Reticulum / Q000378:metabolism; D005372:Filipin; D047908:Intracellular Signaling Peptides and Proteins; D008562:Membrane Glycoproteins; D008856:Microscopy, Fluorescence; D008954:Models, Biological; D008969:Molecular Sequence Data; D009154:Mutation; D000077296:Niemann-Pick C1 Protein; D009542:Niemann-Pick Diseases / Q000378:metabolism*; D011506:Proteins / Q000378:metabolism*; D014162:Transfection
chemical_list: D002352:Carrier Proteins; D008078:Cholesterol, LDL; D047908:Intracellular Signaling Peptides and Proteins; D008562:Membrane Glycoproteins; C106881:NPC1 protein, human; D000077296:Niemann-Pick C1 Protein; D011506:Proteins; D005372:Filipin