Bru-Martínez Roque, Martínez-Márquez Ascensión, Morante-Carriel Jaime, Sellés-Marchart Susana, Martínez-Esteso María José, Pineda-Lucas José Luis, Luque Ignacio
Plant Proteomics and Functional Genomics Group, Department of Agrochemistry and Biochemistry, Faculty of Science and IMEM Ramon Margalef, University of Alicante, Ctra. San Vicente del Raspeig s/n, 03080, Alicante, Spain.
Biotechnology and Molecular Biology Group, Quevedo State Technical University, Quevedo, 120501, Ecuador.
Methods Mol Biol. 2018;1696:147-162. doi: 10.1007/978-1-4939-7411-5_10.
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.
诸如选择反应监测/多反应监测（SRM/MRM）等靶向质谱方法已在蛋白质检测和定量中得到广泛应用，可与传统免疫亲和技术相媲美。它提供了一种通用程序，用于基于对通常由胰蛋白酶消化产生的替代肽段的直接分析，开发一种快速、高特异性、灵敏、准确且廉价的蛋白质靶向检测和定量方法。该方法可有利地应用于植物蛋白质组学领域，特别是对于非模式物种，因为几乎没有免疫试剂可用。在此，我们描述了为开发一种MRM方法以检测和定量来自非模式且数据库中代表性不足的物种枇杷（Eriobotrya japonica Lindl.）的类囊体结合蛋白多酚氧化酶的同工型而需要考虑的问题。
