1. Physiological and proteomic approaches to evaluate the role of sterol binding in elicitin-induced resistance
- Author
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Michal Obořil, Nikola Ptáčková, Ladislav Dokládal, Tomáš Kašparovský, Sylvain Jeandroz, Jiří Damborský, Markéta Žd'árská, Radka Chaloupková, Jan Lochman, Zbyněk Zdráhal, Karel Stejskal, University of science (Masaryk University), Masaryk University, Agroécologie [Dijon], Institut National de la Recherche Agronomique (INRA)-Université de Bourgogne (UB)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement, Faculty of Science [Brno] (SCI / MUNI), Masaryk University [Brno] (MUNI), Loschmidt Laboratories, Department of Experimental Biology, Faculty of Science, Institut National de la Recherche Agronomique ( INRA ) -Université de Bourgogne ( UB ) -AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Université Bourgogne Franche-Comté ( UBFC ), and Department of biochemistry, Masaryk University
- Subjects
Phytophthora ,Proteomics ,0106 biological sciences ,Physiology ,Mutant ,Phospholipid ,Plant Science ,Biology ,omega-loop conformation ,01 natural sciences ,Fungal Proteins ,Structure-Activity Relationship ,03 medical and health sciences ,chemistry.chemical_compound ,Protein structure ,[ SDV.SA.AGRO ] Life Sciences [q-bio]/Agricultural sciences/Agronomy ,Tobacco ,[SDV.BV]Life Sciences [q-bio]/Vegetal Biology ,Plant Immunity ,intercellular proteome ,Phospholipids ,Plant Diseases ,030304 developmental biology ,0303 health sciences ,Phytophthora cryptogea ,Proteins ,Elicitin ,biology.organism_classification ,Research Papers ,Recombinant Proteins ,Cell biology ,Elicitor ,Plant Leaves ,sterol binding ,Sterols ,resistance induction ,Biochemistry ,chemistry ,ω-loop conformation ,Mutation ,Proteome ,Elicitins ,Sterol binding ,Chlorogenic Acid ,Reactive Oxygen Species ,Sesquiterpenes ,Protein Binding ,010606 plant biology & botany - Abstract
International audience; Cryptogein is a proteinaceous elicitor secreted by Phytophthora cryptogea that can induce resistance to P. parasitica in tobacco plants. On the basis of previous computer modelling experiments, by site-directed mutagenesis a series of cryptogein variants was prepared with altered abilities to bind sterols, phospholipids or both. The sterol binding and phospholipid transfer activities corresponded well with the previously reported structural data. Induction of the synthesis of reactive oxygen species (ROS) in tobacco cells in suspension and proteomic analysis of intercellular fluid changes in tobacco leaves triggered by these mutant proteins were not proportional to their ability to bind or transfer sterols and phospholipids. However, changes in the intercellular proteome corresponded to transcription levels of defence genes and resistance to P. parasitica and structure-prediction of mutants did not reveal any significant changes in protein structure. These results suggest, contrary to previous proposals, that the sterol-binding ability of cryptogein and its mutants, and the associated conformational change in the omega-loop, might not be principal factors in either ROS production or resistance induction. Nevertheless, the results support the importance of the omega-loop for the interaction of the protein with the high affinity binding site on the plasma membrane.
- Published
- 2012
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