title: Mediation of elicitin activity on tobacco is assumed by elicitin-sterol complexes.
abstract: Elicitins secreted by phytopathogenic Phytophthora spp. are proteinaceous elicitors of plant defense mechanisms and were demonstrated to load, carry, and transfer sterols between membranes. The link between elicitor and sterol-loading properties was assessed with the use of site-directed mutagenesis of the 47 and 87 cryptogein tyrosine residues, postulated to be involved in sterol binding. Mutated cryptogeins were tested for their ability to load sterols, bind to plasma membrane putative receptors, and trigger biological responses. For each mutated elicitin, the chemical characterization of the corresponding complexes with stigmasterol (1:1 stoichiometry) demonstrated their full functionality. However, these proteins were strongly altered in their sterol-loading efficiency, specific binding to high-affinity sites, and activities on tobacco cells. Ligand replacement experiments strongly suggest that the formation of a sterol-elicitin complex is a requisite step before elicitins fasten to specific binding sites. This was confirmed with the use of two sterol-preloaded elicitins. Both more rapidly displaced labeled cryptogein from its specific binding sites than the unloaded proteins. Moreover, the binding kinetics of elicitins are related to their biological effects, which constitutes the first evidence that binding sites could be the biological receptors. The first event involved in elicitin-mediated cell responses is proposed to be the protein loading with a sterol molecule.
authors.affiliation: Unité Mixte de Recherche 692, Laboratoire de Phytopharmacie et de Biochimie des Interactions Cellulaires, Institut National de la Recherche Agronomique, 21065 Dijon-cedex, France.
authors.forename: H
authors.initials: H
authors.lastname: Osman
authors.forename: S
authors.initials: S
authors.lastname: Vauthrin
authors.forename: V
authors.initials: V
authors.lastname: Mikes
authors.forename: M L
authors.initials: ML
authors.lastname: Milat
authors.forename: F
authors.initials: F
authors.lastname: Panabières
authors.forename: A
authors.initials: A
authors.lastname: Marais
authors.forename: S
authors.initials: S
authors.lastname: Brunie
authors.forename: B
authors.initials: B
authors.lastname: Maume
authors.forename: M
authors.initials: M
authors.lastname: Ponchet
authors.forename: J P
authors.initials: JP
authors.lastname: Blein
mesh_terms: D020418:Algal Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology*; D001665:Binding Sites; D002118:Calcium / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D005656:Fungal Proteins; D006790:Host-Parasite Interactions; D006863:Hydrogen-Ion Concentration; D008954:Models, Biological; D008958:Models, Molecular; D010838:Phytophthora / Q000502:physiology; D010935:Plant Diseases / Q000469:parasitology; D010940:Plant Proteins / Q000378:metabolism; D011485:Protein Binding; D011487:Protein Conformation; D020033:Protein Isoforms; D011506:Proteins; D011956:Receptors, Cell Surface / Q000378:metabolism; D013261:Sterols / Q000378:metabolism*; D013997:Time Factors; D014026:Nicotiana / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism* / Q000469:parasitology; D014443:Tyrosine / Q000235:genetics / Q000378:metabolism
chemical_list: D020418:Algal Proteins; D005656:Fungal Proteins; D010940:Plant Proteins; D020033:Protein Isoforms; D011506:Proteins; D011956:Receptors, Cell Surface; D013261:Sterols; C061032:cryptogein protein, Phytophthora cryptogea; C085101:elicitin, Phytophthora; D014443:Tyrosine; D002118:Calcium