iTRAQ鸟枪法蛋白质组学在精神疾病研究中用于测量脑蛋白的应用。

Application of iTRAQ Shotgun Proteomics for Measurement of Brain Proteins in Studies of Psychiatric Disorders.

作者信息

Núñez Erika Velásquez, Guest Paul C, Martins-de-Souza Daniel, Domont Gilberto Barbosa, Nogueira Fábio César Sousa

机构信息

Laboratory of Protein Chemistry - Proteomics Unit, Chemistry Institute, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil.

Department of Biochemistry and Tissue Biology, Laboratory of Neuroproteomics, Institute of Biology, University of Campinas, CidadeUniversitáriaZeferinoVaz, Campinas, Brazil.

出版信息

Adv Exp Med Biol. 2017;974:219-227. doi: 10.1007/978-3-319-52479-5_18.

DOI:10.1007/978-3-319-52479-5_18

PMID:28353239

Abstract

Shotgun proteomics has been used successfully for more than two decades for measurement of proteins from diverse biological systems. This has led to new insights and identification of potential biomarkers and drug targets for numerous medical conditions. The advent of mass-labeling approaches such as isobaric tagging for relative and absolute quantitation (iTRAQ) has allowed multiplexing of samples to improve the accuracy and throughput of experiments and the reduction of costs. Here, we describe a detailed iTRAQ mass spectrometry protocol for simultaneous analysis of four proteomes from postmortem brain tissue extracts. In addition, a post-labeling procedure for peptide fractionation is presented.

摘要

鸟枪法蛋白质组学已成功应用二十多年，用于测量来自不同生物系统的蛋白质。这带来了新的见解，并为众多医学病症鉴定了潜在的生物标志物和药物靶点。诸如用于相对和绝对定量的等压标记（iTRAQ）等质量标记方法的出现，使得能够对样品进行多重分析，以提高实验的准确性和通量，并降低成本。在此，我们描述了一种详细的iTRAQ质谱分析方案，用于同时分析死后脑组织提取物中的四个蛋白质组。此外，还介绍了一种用于肽段分级分离的标记后程序。