title: Isolation of Protein Storage Vacuoles and Their Membranes.
abstract: Protein-storage vacuoles (PSVs) are specialized vacuoles that sequester large amounts of storage proteins. During seed development, PSVs are formed de novo and/or from preexisting lytic vacuoles. Seed PSVs can be subdivided into four distinct compartments: membrane, globoid, matrix, and crystalloid. In this chapter, we introduce easy methods for isolation of PSVs and their membranes from pumpkin seeds. These methods facilitate the identification and characterization of PSV components.
authors.affiliation: Department of Botany, Graduate School of Science, Kyoto University, Kitashirakawa, Sakyoku, Kyoto, 606-8502, Japan.
authors.forename: Tomoo
authors.initials: T
authors.lastname: Shimada
authors.affiliation: Department of Botany, Graduate School of Science, Kyoto University, Kitashirakawa, Sakyoku, Kyoto, 606-8502, Japan. ihnishi@gr.bot.kyoto-u.ac.jp.
authors.forename: Ikuko
authors.initials: I
authors.lastname: Hara-Nishimura
mesh_terms: D002458:Cell Fractionation / Q000295:instrumentation / Q000379:methods*; D002499:Centrifugation, Density Gradient / Q000295:instrumentation / Q000379:methods; D028464:Cucurbita / Q000737:chemistry*; D003470:Culture Media / Q000737:chemistry; D007425:Intracellular Membranes / Q000737:chemistry* / Q000648:ultrastructure; D008853:Microscopy; D010940:Plant Proteins / Q000737:chemistry; D011205:Povidone / Q000737:chemistry; D021381:Protein Transport; D055314:Seed Storage Proteins / Q000737:chemistry*; D012639:Seeds / Q000737:chemistry*; D012822:Silicon Dioxide / Q000737:chemistry; D014617:Vacuoles / Q000737:chemistry*
chemical_list: D003470:Culture Media; D010940:Plant Proteins; D055314:Seed Storage Proteins; C016039:Percoll; D012822:Silicon Dioxide; D011205:Povidone
keywords: Cucurbita maxima; Protein-storage vacuole; Pumpkin seed; Storage protein; Tonoplast; Vacuole