title: Purification and identification of a Ca(2+)-pectate binding peroxidase from Arabidopsis leaves.
abstract: A protein fraction was obtained from Arabidopsis (Arabidopsis thaliana, L.) leaf extract by affinity chromatography through a Ca(2+)-pectate/polyacrylamide gel. Further purification by preparative isoelectric focusing and SDS PAGE allowed the separation of a peroxidase that was identified as being peroxidase AtPrx34 (AtprxCb, accession number X71794) by N-terminal amino acid microsequencing. AtPrx34 belongs to a group of five Arabidopsis sequences encoding putative pectin-binding peroxidases. An expression study showed that it is expressed in root, stem, flower and leaf. It was produced by Escherichia coli and tested for its ability to bind to Ca(2+)-pectate. The identity of the amino acids involved in the interaction between the peroxidase and the Ca(2+)-pectate structure is discussed.
authors.affiliation: Laboratoire de Physiologie Végétale, Université de Genève, Quai Ernest-Ansermet 30, CH-1211 Genève 4, Switzerland.
authors.forename: Kavita
authors.initials: K
authors.lastname: Shah
authors.forename: Claude
authors.initials: C
authors.lastname: Penel
authors.forename: Jean
authors.initials: J
authors.lastname: Gagnon
authors.forename: Christophe
authors.initials: C
authors.lastname: Dunand
mesh_terms: D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000201:enzymology*; D029681:Arabidopsis Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D001665:Binding Sites; D015152:Blotting, Northern; D002135:Calcium-Binding Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D007525:Isoelectric Focusing; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010368:Pectins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D010544:Peroxidases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D018515:Plant Leaves / Q000201:enzymology; D018514:Plant Structures / Q000378:metabolism
chemical_list: D029681:Arabidopsis Proteins; D002135:Calcium-Binding Proteins; D010368:Pectins; D010544:Peroxidases