title: Light induces rapid changes of the phosphorylation pattern in the cytosol of evacuolated parsley protoplasts.
abstract: The fractionation of cells of a parsley suspension culture [Petroselinum crispum (Mill.) A. Hill] by protoplasting and subsequent removal of the vacuoles led to physiologically intact evacuolated protoplasts retaining light inducibility of chalcone synthase expression. Lysis of the evacuolated protoplasts permitted the isolation of a pure, highly concentrated cytosolic fraction containing major cytosolic membranes but only minor contamination by proplastids, mitochondria, and nuclei. Short-time irradiations of the cytosol with red or UV-containing white light resulted in very fast changes of the phosphorylation pattern of 18-, 40-, 48-, 55- to 70-, and 120-kDa proteins. Major differences were observed between the phosphorylation patterns obtained by red or UV-containing white light treatment, indicating a different primary action of the excited photoreceptors in vitro. Separation of the microsomal fraction from the cytosolic matrix established the localization of these proteins. Chase and photoreversibility experiments revealed that phytochrome in vitro regulates the phosphorylation of the 40-kDa protein by modifying a soluble cytosolic kinase/phosphatase system.
authors.affiliation: Department of Biology II/Botany, University of Freiburg, Federal Republic of Germany.
authors.forename: K
authors.initials: K
authors.lastname: Harter
authors.forename: H
authors.initials: H
authors.lastname: Frohnmeyer
authors.forename: S
authors.initials: S
authors.lastname: Kircher
authors.forename: T
authors.initials: T
authors.lastname: Kunkel
authors.forename: S
authors.initials: S
authors.lastname: Mühlbauer
authors.forename: E
authors.initials: E
authors.lastname: Schäfer
mesh_terms: D002458:Cell Fractionation; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D003600:Cytosol / Q000378:metabolism / Q000528:radiation effects; D015151:Immunoblotting; D008027:Light; D010766:Phosphorylation / Q000528:radiation effects; D010940:Plant Proteins / Q000378:metabolism; D010944:Plants / Q000378:metabolism / Q000528:radiation effects* / Q000648:ultrastructure; D011523:Protoplasts; D015398:Signal Transduction / Q000528:radiation effects; D014617:Vacuoles
chemical_list: D010940:Plant Proteins