Targeted Quantification of Isoforms of a Thylakoid-Bound Protein: MRM Method Development

• DOI: 10.1007/978-1-4939-7411-5_10
• Publication Date: 2017-10-30T01:44:22Z
• Authors: Roque Bru-Martínez; Ascensión Martíne...; Jaime Morante-Car...; Susana Sellés-Mar...; María José Martín...; José Luis Pineda-...; Ignacio Luque
• Keywords: Proteomics; Chloroplast Proteins; Eriobotrya; Membrane Proteins; Protein Isoforms; Thylakoids; Chromatography, High Pressure Liquid; Mass Spectrometry

Targeted mass spectrometric methods such as selected/multiple reaction monitoring (SRM/MRM) have found intense application in protein detection and quantification which competes with classical immunoaffinity techniques. It provides a universal procedure to develop a fast, highly specific, sensitive, accurate, and cheap methodology for targeted detection and quantification of proteins based on the direct analysis of their surrogate peptides typically generated by tryptic digestion. This methodology can be advantageously applied in the field of plant proteomics and particularly for non-model species since immunoreagents are scarcely available. Here, we describe the issues to take into consideration in order to develop a MRM method to detect and quantify isoforms of the thylakoid-bound protein polyphenol oxidase from the non-model and database underrepresented species Eriobotrya japonica Lindl.