Differential protein profiling of synovial fluid from rheumatoid arthritis and osteoarthritis patients using LC-MALDI TOF/TOF
Differential protein profiling of synovial fluid from rheumatoid arthritis and osteoarthritis patients using LC-MALDI TOF/TOF
复制标题
DOI：
10.1016/j.jprot.2011.12.042
复制
发表时间：
2012-06-06
影响因子：
3.3
通讯作者：
Blanco, Francisco J.
中科院分区：
文献类型：
--
作者：
Mateos, Jesus;Lourido, Lucia;Blanco, Francisco J.
本研究的目的是确定这些蛋白质相对更丰富的滑液（SF）的患者患有类风湿性关节炎（RA）和骨关节炎（OA），使用高效液相色谱-质谱联用。将来自每种疾病的20份个体SF样本合并为两组（RA和OA），以减少极端个体值的贡献。在蛋白质组学分析之前，将样品从前20种最丰富的血浆蛋白中免疫耗竭，以富集较低丰度的蛋白质级分。然后，它们进行蛋白质大小分级和凝胶内消化，然后在纳米LC系统中进行反相肽分离，随后通过MALDI-TOF/TOF进行肽鉴定。结果表明，在SF中共鉴定出136种蛋白质，是迄今为止蛋白质组学研究最多的SF蛋白质，并通过光谱计数法对RA和OA的蛋白质进行了相对定量。在RA中，我们的研究结果显示与补体激活、炎症和免疫反应相关的蛋白质的相对丰度更高，例如主要的基质金属蛋白酶和几种嗜中性粒细胞相关蛋白质。在OA中，我们检测到参与细胞外基质（ECM）形成和重塑的蛋白质增加，如纤连蛋白、激肽原-1、软骨酸性蛋白1和软骨寡聚基质蛋白。通过免疫印迹分析验证了MMP-1、BGH 3、纤连蛋白和凝溶胶蛋白的结果。在这项工作中鉴定的一些新蛋白质可能不仅与增加RA和OA过程的发病机制的知识有关，而且还作为推定的疾病生物标志物，因为它们在SF中的存在是它们在血清中稀释的前一步。本文是特刊的一部分，题为：蛋白质组学：临床联系。(C)2012爱思唯尔有限公司版权所有。
The purpose of this study was to identify those proteins relatively more abundant in the synovial fluid (SF) of patients suffering from rheumatoid arthritis (RA) and osteoarthritis (OA) using high performance liquid chromatography coupled to mass spectrometry. 20 individual SF samples from each disease were pooled into two groups (RA and OA) to reduce the contribution of extreme individual values. Prior to the proteomic analysis, samples were immunodepleted from the top 20 most abundant plasma proteins, to enrich the lower-abundance protein fractions. Then, they were subjected to protein size fractioning and in-gel digestion, followed by reversed-phase peptide separation in a nano-LC system and subsequent peptide identification by MALDI-TOF/TOF. This strategy led to the identification of 136 different proteins in SF, which is the largest number of SF proteins described up to date by proteomics.A relative quantification of the proteins between RA and OA was carried out by spectral counting analysis. In RA, our results show a greater relative abundance of proteins related to complement activation, inflammation and the immune response, such as the major matrix metalloproteinases and several neutrophil-related proteins. In OA, we detected an increase in proteins involved in the formation and remodeling of the extracellular matrix (ECM), such as fibronectin, kininogen-1, cartilage acidic protein 1 and cartilage oligomeric matrix protein. The results obtained for MMP-1, BGH3, fibronectin and gelsolin were verified by immunoblotting analyses. Some of the novel proteins identified in this work might be relevant not only for increasing knowledge on the etiopathogenesis of RA and OA processes, but also as putative disease biomarkers, as their presence in SF is a prior step to their dilution in serum. This article is part of a Special Issue entitled: Proteomics: The clinical link. (C) 2012 Elsevier B.V. All rights reserved.