title: Enrichment of the Plant Cytosolic Fraction.
abstract: The cytosol is at the core of cellular metabolism and contains many important metabolic pathways, including glycolysis, gluconeogenesis, and the pentose phosphate pathway. Despite the importance of this matrix, few attempts have sought to specifically enrich this compartment from plants. Although a variety of biochemical pathways and signaling cascades pass through the cytosol, much of the focus has usually been targeted at the reactions that occur within membrane-bound organelles of the plant cell. In this chapter, we outline a method for the enrichment of the cytosol from rice suspension cell cultures which includes sample preparation and enrichment as well as validation using immunoblotting and fluorescence-tagged proteins.
authors.affiliation: Joint BioEnergy Institute and Biological Systems and Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, CA, 94702, USA.
authors.forename: Jeemeng
authors.initials: J
authors.lastname: Lao
authors.affiliation: Joint BioEnergy Institute and Biological Systems and Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, CA, 94702, USA.
authors.forename: Andreia M
authors.initials: AM
authors.lastname: Smith-Moritz
authors.affiliation: Joint BioEnergy Institute and Biological Systems and Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, CA, 94702, USA.
authors.forename: Jennifer C
authors.initials: JC
authors.lastname: Mortimer
authors.affiliation: Joint BioEnergy Institute and Biological Systems and Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, CA, 94702, USA. jheazlewood@unimelb.edu.au.|School of BioSciences, The University of Melbourne, Melbourne, VIC, 3010, Australia. jheazlewood@unimelb.edu.au.
authors.forename: Joshua L
authors.initials: JL
authors.lastname: Heazlewood
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D015415:Biomarkers / Q000378:metabolism; D015153:Blotting, Western; D018929:Cell Culture Techniques; D002458:Cell Fractionation / Q000295:instrumentation / Q000379:methods*; D003470:Culture Media / Q000737:chemistry; D003600:Cytosol / Q000737:chemistry* / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D015870:Gene Expression; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D013058:Mass Spectrometry; D019697:Onions / Q000737:chemistry* / Q000254:growth & development; D012275:Oryza / Q000737:chemistry* / Q000254:growth & development; D059828:Plant Cells / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure; D010940:Plant Proteins / Q000235:genetics* / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D015415:Biomarkers; D003470:Culture Media; D008164:Luminescent Proteins; D010940:Plant Proteins; C527701:enhanced cyan fluorescent protein; C054562:yellow fluorescent protein, Bacteria; D049452:Green Fluorescent Proteins
keywords: Cell culture; Cytoplasm; Cytosol; Rice