title: Reconstitution of Phospholipase A2-Dependent Golgi Membrane Tubules.
abstract: The Golgi complex is the Grand Central Station of intracellular membrane trafficking in the secretory and endocytic pathways. Anterograde and retrograde export of cargo from the Golgi complex involves a complex interplay between the formation of coated vesicles and membrane tubules, although much less is known about tubule-mediated trafficking. Recent advances using in vitro assays have identified several cytoplasmic phospholipase A2 (PLA2) enzymes that are required for the biogenesis of membrane tubules and their roles in the functional organization of the Golgi complex. In this chapter we describe methods for the cell-free reconstitution of PLA2-dependent Golgi membrane tubule formation. These methods should facilitate the identification of other proteins that regulate this process.
authors.affiliation: Department of Biology, Ithaca College, Ithaca, NY, 14850, USA.
authors.forename: Edward B
authors.initials: EB
authors.lastname: Cluett
authors.affiliation: Department of Microbial Pathogenesis and Immunology, Texas A&M Health Science Center, Bryan, TX, 77807, USA.|Department of Veterinary Pathobiology, Texas A&M University, College Station, TX, 77843, USA.|Norman Borlaug Center, Texas A&M University, College Station, TX, 77843, USA.
authors.forename: Paul
authors.initials: P
authors.lastname: de Figueiredo
authors.affiliation: MRC Centre for Regenerative Medicine, The University of Edinburgh, 5 Little France Drive, Edinburgh, EH16 4UU, UK.
authors.forename: Marie E
authors.initials: ME
authors.lastname: Bechler
authors.affiliation: Department of Molecular Biology and Genetics, Cornell University, Ithaca, NY, 14850, USA.
authors.forename: Kevin D
authors.initials: KD
authors.lastname: Thorsen
authors.affiliation: Department of Molecular Biology and Genetics, Cornell University, Ithaca, NY, 14850, USA. wjb5@cornell.edu.
authors.forename: William J
authors.initials: WJ
authors.lastname: Brown
mesh_terms: D000818:Animals; D006056:Golgi Apparatus / Q000201:enzymology*; D006801:Humans; D007425:Intracellular Membranes / Q000201:enzymology*; D008099:Liver / Q000201:enzymology*; D054467:Phospholipases A2 / Q000737:chemistry / Q000378:metabolism; D051381:Rats
chemical_list: D054467:Phospholipases A2
keywords: Golgi complex; In vitro reconstitution; Membrane tubules; PLA2 enzymes