Reconstitution in vitro of the components and conditions required for the oxidative cross-linking of extracellular proteins in French bean (Phaseolus vulgaris L.).
Oxidative cross linking of three (hydroxy)proline-rich cell wall proteins, known to be immobilised during the elicitor-induced oxidative burst in French bean cells, was modelled using peroxidases with cysteine or H2O2. Further reconstitution of the homologous system was achieved with a 46 kDa bean cell-wall peroxidase using conditions known to appertain in cell walls prior to the immobilisation. Thus, cell wall alkalinisation and secretion of a reductant have been reconstituted by addition of apoplastic fluid to the wall proteins and the 46 kDa peroxidase with resultant cross-linking. This is the first demonstration of the oxidative cross-linking of cell wall glycoproteins without the addition of external H2O2.
Wojtaszek
P
P
Division of Biochemistry, School of Biological Sciences, Royal Holloway, University of London, Egham, Surrey, UK.
Trethowan
J
J
Bolwell
G P
GP
D003432:Cross-Linking Reagents / Q000737:chemistry / Q000494:pharmacology*; D003545:Cysteine / Q000494:pharmacology; D007887:Fabaceae; D006023:Glycoproteins / Q000737:chemistry / Q000187:drug effects; D006735:Horseradish Peroxidase / Q000494:pharmacology; D006861:Hydrogen Peroxide / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D010084:Oxidation-Reduction; D010940:Plant Proteins / Q000737:chemistry* / Q000187:drug effects; D010946:Plants, Medicinal
D003432:Cross-Linking Reagents; D006023:Glycoproteins; D010940:Plant Proteins; C005488:extensin protein, plant; D006861:Hydrogen Peroxide; D006735:Horseradish Peroxidase; D003545:Cysteine