Absolute protein quantification by LC/MS(E) for global analysis of salicylic acid-induced plant protein secretion responses

The plant cell wall is a dynamic cellular compartment consisting of a complex matrix of components that can change dramatically in response to environmental stresses. During pathogen attack, for instance, a wide spectrum of proteins that participate in various sequential processes involved in plant...

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Bibliographic Details
Published in:Journal of proteome research 2009-01, Vol.8 (1), p.82-93
Main Authors: Cheng, Fang-yi, Blackburn, Kevin, Lin, Yu-min, Goshe, Micheal B, Williamson, John D
Format: Article
Language:English
Subjects:
Arabidopsis - drug effects
Arabidopsis - metabolism
Chromatography, Liquid - methods
Computational Biology
Mass Spectrometry - methods
Plant Physiological Phenomena
Plant Proteins - metabolism
Plant Proteins - secretion
Proteins - secretion
Proteome
Proteomics - methods
Salicylic Acid - metabolism
Salicylic Acid - pharmacology
Time Factors
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Summary:The plant cell wall is a dynamic cellular compartment consisting of a complex matrix of components that can change dramatically in response to environmental stresses. During pathogen attack, for instance, a wide spectrum of proteins that participate in various sequential processes involved in plant defense is secreted into the cell wall. In this study, a mass spectrometry, data-independent acquisition approach known as LC/MS (E) was used to assess temporal changes in the cell wall proteome in response to different levels of an endogenous inducer of plant disease defense responses, salicylic acid (SA). LC/MS (E) was used as a label-free method that enabled simultaneous protein identification and absolute femtomole quantification of each protein secreted into the extracellular matrix. A total of 74 secreted proteins were identified, 63 of which showed increased specific secretion in response to SA. A majority of this induced secretion occurred within 2 h of treatment, indicating that many proteins are involved in the early stages of plant defenses. We also identified a number of apparently nonclassically secreted proteins, suggesting that, as in many nonplant systems, Golgi/ER-independent mechanisms exist for plant protein secretion. These results provide new insight into plant apoplastic defense mechanisms and demonstrate that LC/MS (E) is a powerful tool for obtaining both relative and absolute proteome-scale quantification that can be applied to complex, time- and dose-dependent experimental designs.
ISSN:1535-3893
DOI:10.1021/pr800649s