title: Differential plant proteome analysis by isobaric tags for relative and absolute quantitation (iTRAQ).
abstract: Protein relative quantitation is one of the main targets in many proteomic experiments. Among the range of techniques available for both top-down and bottom-up approaches, isobaric tags for relative and absolute quantitation (iTRAQ) have gained positions within the top-rank techniques used for this purpose in the recent years. Briefly, each iTRAQ reagent consists of three different components: a reporter group (with a variable mass in the range of 114-117 amu), a balance group, and an amino-reactive group. The isobaric nature of iTRAQ-labeled peptides adds a signal to every peptide in the sample which is detectable in both MS and MS/MS spectra, thus enhancing the sensitivity of detection. During MS/MS, the reporter groups are released as singly charged ions with m/z ratios ranking from 114 to 117 amu, visible in the low mass region of MS/MS spectra. The iTRAQ technology can be used to analyze up to four different samples using the 4-plex kit (reporter groups 114-115 amu) or can be scaled up to eight different samples using the 8-plex kit (reporter groups 113-121 amu). In this chapter, we focus on the experimental procedures typically using 4-plex labeling, including tips leading to successful application of iTRAQ technology for the analysis of plant protein mixtures.
authors.affiliation: Plant Proteomics and Functional Genomics Group, Department of Agrochemistry and Biochemistry, Faculty of Science, University of Alicante, Alicante, Spain.
authors.forename: María J
authors.initials: MJ
authors.lastname: Martínez-Esteso
authors.forename: Juan
authors.initials: J
authors.lastname: Casado-Vela
authors.forename: Susana
authors.initials: S
authors.lastname: Sellés-Marchart
authors.forename: María A
authors.initials: MA
authors.lastname: Pedreño
authors.forename: Roque
authors.initials: R
authors.lastname: Bru-Martínez
mesh_terms: D005591:Chemical Fractionation; D007553:Isotope Labeling / Q000379:methods*; D013058:Mass Spectrometry; D010940:Plant Proteins / Q000378:metabolism*; D020543:Proteome / Q000378:metabolism*; D013223:Statistics as Topic; D027843:Vitis / Q000378:metabolism*
chemical_list: D010940:Plant Proteins; D020543:Proteome