title: Purification of a protein phosphatase from chloroplast stroma capable of dephosphorylating the light-harvesting complex-II.
abstract: A protein phosphatase was purified from the stroma of Pea (Pisum sativum L.) chloroplasts that is capable of dephosphorylating synthetic phosphopeptides. Following chromatographic purification of greater than 400-fold, two-dimensional electrophoresis indicated that the stromal protein phosphatase is a 29-kD protein. A similar molecular size was determined for the protein-phosphatase activity using gel-permeation chromatography, indicating that the stromal protein phosphatase is probably a monomer. The purified enzyme was able to dephosphorylate synthetic phosphopeptides, which mimic the thylakoid light-harvesting complex II (LHC-II) N terminus, as well as LHC-II in thylakoid membranes, but did not dephosphorylate the major 64-kD phosphoprotein in the stroma. The stromal protein phosphatase did not discriminate between dephosphorylation of phosphothreonine and phosphoserine residues in synthetic peptide substrates, providing further evidence that this enzyme is distinct from the protein phosphatase localized in thylakoid membranes. The exact physiological role of the stromal protein phosphatase has yet to be determined, but it may function in the dephosphorylation of LHC-II.
authors.affiliation: Department of Biochemistry, University of Nebraska, Lincoln 68588-0664, USA.
authors.forename: M F
authors.initials: MF
authors.lastname: Hammer
authors.forename: J
authors.initials: J
authors.lastname: Markwell
authors.forename: G
authors.initials: G
authors.lastname: Sarath
mesh_terms: D000595:Amino Acid Sequence; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D008969:Molecular Sequence Data; D018532:Pisum sativum / Q000201:enzymology; D010749:Phosphoprotein Phosphatases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D010766:Phosphorylation; D045322:Photosynthetic Reaction Center Complex Proteins / Q000378:metabolism*
chemical_list: D045322:Photosynthetic Reaction Center Complex Proteins; D010749:Phosphoprotein Phosphatases