Protein Identification and Quantification by Mass Spectrometry-Based Analysis: Applications in Plant-Pathogen Interactions Studies

Carmen      Garcia-Limones; Jesus      Mercado-Blanco; Inmaculada      Jorge

doi:10.2174/157016410793611738

Abstract

Mass spectrometry (MS) has become an essential technology for proteomics applications in biological sciences. Advances in this technique have been possible owing to improvements in MS instrumentation, new experimental strategies in sample preparation, and development of bioinformatics tools for data analyses. In recent years, complementary strategies to the classical two-dimensional gel electrophoresis approaches (2-DE) have been developed. These techniques are based on multidimensional peptide separation coupled to tandem MS (also referred as “second generation proteomics”), enabling protein expression analysis and high throughput protein identification studies. New methods such as Multidimensional Protein Identification Technology (MudPIT) and stable isotope labeling of protein/peptide samples (either by chemical, metabolic, or enzymatic methods), among others, are powerful tools for large-scale studies on characterization and expression of proteins in complex biological systems. Hence, these techniques can be very useful in the study of plant-pathogen interactions, aiding to detect and characterize both plant proteins concerned in defense reactions and pathogen proteins involved in pathogenicity and/or virulence. But these techniques have been implemented in these biological systems just recently. We will examine here how MS-based proteomics approaches are helping to better understand the multifaceted phenomena underlying plant-pathogen interactions.

Keywords: Mass spectrometry, proteomics, plant-pathogen interaction, protein identification, protein quantification, omics cascade, second generation proteomics, peptide level proteomics, effector-triggered immunity, hypersensitive response, pathogenassociated molecular patterns, Oryza sativa, Magnaporthe grisea, Fusarium graminearum, Triticum aestivum, MudPIT Multidimensional, Protein Identification Technology, Arabidopsis thaliana, Phaseolus vulgaris, fungal xylanase, Detergent-Resistant Membranes (DRMs), Phytophthora, Isotope-Coded Affinity Tags, ICAT, isobaric Tag for Relative and Absolute Quantification, iTRAQ, heat shock protein, HSP81, plastid associated protein, PAP, fibrillin, NADPH oxidase, Picea abies