title: Analysis of protein transport to lysosomes.
abstract: Lysosomes are terminal degradative organelles that are found in all higher eukaryotic cells. The biogenesis of lysosomes involves the transport of various acid hydrolases and transmembrane glycoproteins from their site of synthesis in the endoplasmic reticulum through the biosynthetic and endocytic pathways. Protein transport to lysosomes can be studied by a combination of techniques based on the separation of intracellular organelles. Percoll density gradient centrifugation has long been the method of choice for separating lysosomes from other organelles in cell homogenates, and accordingly, this unit describes protocols for obtaining reasonably pure lysosomal fractions from mammalian cells using Percoll density gradient separation.
authors.affiliation: University of Zurich, Zurich, Switzerland.
authors.forename: Beat E
authors.initials: BE
authors.lastname: Schaub
authors.affiliation: University of Zurich, Zurich, Switzerland.
authors.forename: Prashant
authors.initials: P
authors.lastname: Nair
authors.affiliation: University of Zurich, Zurich, Switzerland.
authors.forename: Jack
authors.initials: J
authors.lastname: Rohrer
mesh_terms: D000818:Animals; D001671:Biochemistry / Q000379:methods*; D018929:Cell Culture Techniques; D002458:Cell Fractionation / Q000379:methods*; D002460:Cell Line; D002499:Centrifugation, Density Gradient / Q000379:methods; D006801:Humans; D008247:Lysosomes / Q000378:metabolism*; D051379:Mice; D008967:Molecular Biology / Q000379:methods*; D011205:Povidone / Q000737:chemistry; D021381:Protein Transport / Q000502:physiology; D012822:Silicon Dioxide / Q000737:chemistry; D013347:Subcellular Fractions / Q000378:metabolism
chemical_list: C016039:Percoll; D012822:Silicon Dioxide; D011205:Povidone