期刊
JOURNAL OF PROTEOME RESEARCH
卷 8, 期 3, 页码 1168-1182出版社
AMER CHEMICAL SOC
DOI: 10.1021/pr800619h
关键词
Phosphoproteomics; titanium dioxide; label-free quantification; LC-MS/MS; tomato; hypersensitive response; defense signaling; photosynthesis; Hsp90; low-oxygen stress
资金
- Dutch Organization for Scientific Research (NWO) [864.02.008]
- Netherlands Genomics Initiative [050-72-416]
- Lundbeck Foundation Research Professor
- Danish Research Agency
An important mechanism by which plants defend themselves against pathogens is the rapid execution of a hypersensitive response (HR). Tomato plants containing the Cf-4 resistance gene mount an HR that relies on the activation of phosphorylation cascades, when challenged with the Avr4 elicitor secreted by the pathogenic fungus Cladosporium fulvum. Phosphopeptides were isolated from tomato seedlings expressing both Cf-4 and Avr4 using titanium dioxide columns and LC-MS/MS analysis led to the identification of 50 phosphoproteins, most of which have not been described in tomato before. Phosphopeptides were quantified using a label-free approach based on the MS peak areas. We identified 12 phosphopeptides for which the abundance changed upon HR initiation, as compared to control seedlings. Our results suggest that photosynthetic activity is specifically suppressed in a phosphorylation-dependent way during the very-early stages of HR development. In addition, phosphopeptides originating from four Hsp90 isoforms exhibited altered abundances in Cf-4/Avr4 seedlings compared to control seedlings, suggesting that the isoforms of this chaperone protein have a different function in defense signaling. We show that label-free relative quantification of the phosphoproteome of complex samples is feasible, allowing extension of our knowledge on the general physiology and defense signaling of plants mounting the HR.
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