title: Methods for analyzing the role of phospholipase A₂ enzymes in endosome membrane tubule formation.
abstract: Cargo export from mammalian endosomal compartments often involves membrane tubules, into which soluble and membrane-bound cargos are segregated for subsequent intracellular transport. These membrane tubules are highly dynamic and their formation is mediated by a variety of endosome-associated proteins. However, little is known about how these membrane tubules are temporally or spatially regulated, so other tubule-associated proteins are likely to be discovered and analyzed. Therefore, methods to examine the biogenesis and regulation of endosome membrane tubules will prove to be valuable for cell biologists. In this chapter, we describe methods for studying this process using both cell-free, in vitro reconstitution assays, and in vivo image analysis tools.
authors.affiliation: Department of Molecular Biology and Genetics, Cornell University, Ithaca, NY, USA.
authors.forename: Danielle N
authors.initials: DN
authors.lastname: Kalkofen
authors.affiliation: Department of Molecular Biology and Genetics, Cornell University, Ithaca, NY, USA; Department of Microbial Pathogenesis and Immunology, Norman Borlaug Center, Texas A&M University, College Station, TX, USA.
authors.forename: Paul
authors.initials: P
authors.lastname: de Figueiredo
authors.affiliation: Department of Molecular Biology and Genetics, Cornell University, Ithaca, NY, USA.
authors.forename: William J
authors.initials: WJ
authors.lastname: Brown
mesh_terms: D000818:Animals; D001921:Brain / Q000201:enzymology; D002417:Cattle; D011992:Endosomes / Q000201:enzymology / Q000648:ultrastructure*; D006367:HeLa Cells; D006801:Humans; D007425:Intracellular Membranes / Q000201:enzymology / Q000648:ultrastructure*; D051379:Mice; D046529:Microscopy, Electron, Transmission; D054467:Phospholipases A2 / Q000502:physiology*; D021381:Protein Transport; D000067996:RAW 264.7 Cells; D051381:Rats
chemical_list: D054467:Phospholipases A2
keywords: Digital image analysis; Endosome tubules; In vitro reconstitution; Phospholipase A(2)(PLA(2)) enzymes; Transferrin trafficking