Proteomic analysis of human articular cartilage: Identification of differentially expressed proteins in knee osteoarthritis
Proteomic analysis of human articular cartilage: Identification of differentially expressed proteins in knee osteoarthritis
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DOI：
10.1016/j.jbspin.2007.12.003
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发表时间：
2008-07-01
期刊：
影响因子：
4.2
通讯作者：
Zhang, Zhongnan
中科院分区：
文献类型：
--
作者：
Guo, Dunming;Tan, Wenfeng;Zhang, Zhongnan
目的：年龄相关性骨关节炎（OA）的发病机制尚不清楚。为了更好地了解OA的发病机制和关节软骨在OA中进行性破坏的分子基础，我们比较了OA软骨与正常软骨的蛋白质组。方法：去除蛋白多糖和胶原后，提取正常或OA膝关节软骨蛋白质，通过双向凝胶电泳（2-DE）进行分离。结果：正常骨关节炎软骨组织和骨关节炎软骨组织经双向电泳后，分别分离出1436 49个和1472 7个蛋白质点。发现来自OA软骨样品的16个点与正常样品相比在蛋白质量上具有统计学显著的变化。在16个斑点中，LTQ-FT/MS明确地确定了14个蛋白的身份。这些OA相关蛋白分为5组，包括糖酵解和能量产生（ADH，ADK，ENOA，KPYM和FR），信号传导（ANNX-1，PEBP和TUB），氧化还原（PRDX 3和SODM），和软骨基质（COLL-I和COLL-VI）。有趣的是，两个新的RING（真正有趣的新基因）结构域的蛋白质，RF，锌RF，被确定，提出新的途径软骨蛋白regulation.Conclusions：这项研究表明，2-DE其次是LTQ-FT/MS可以成功地用于表征蛋白质组的软骨，而无需在体外培养，可能混淆生理差异。这里描述的独特OA相关蛋白的定义通过证实先前提出的机制和通过定义在疾病发病机制中具有尚未定义的作用的独特参与者来提供对OA的重要机制见解。(c)2008年，Elsevier Masson SAS。All rights reserved.
Objectives: The mechanisms underlying the development of age related osteoarthritis (OA) remain unclear. To better understand the pathogenesis of OA and the molecular basis of progressive destruction of articular cartilage in OA, we compared the proteome of OA cartilage with that of normal cartilage.Methods: After removal of proteoglycans and collagens, proteins extracted from either normal or OA knee joint cartilage were separated by two-dimensional gel electrophoresis (2-DE). The differentially expressed proteins in OA cartilage were chosen to be further identified by linear ion trap-Fourier transform ion cyclotron resonance mass spectrometry (LTQ-FT/MS).Results: A total of 1436 49 or 1472 7 protein spots were resolved by 2-DE of normal or OA cartilage extractions, respectively. Sixteen spots from OA cartilage samples were found to have statistically significant changes in the amount of protein compared with normal samples. Of 16 spots, the identities of 14 proteins were unambiguously determined by LTQ-FT/MS. These OA associated proteins fell into five groups, including glycolysis and energy production (ADH, ADK, ENOA, KPYM and FR), signaling (ANNX-1, PEBP and TUB), Redox (PRDX3 and SODM), and cartilage matrix (COLL-I and COLL-VI). Interestingly, two novel RING (Really Interesting New Gene) domain-containing proteins, RF, Zn-RF, were identified, suggesting novel pathways of cartilage protein regulation.Conclusions: This study shows that 2-DE followed by LTQ-FT/MS can be successfully used to characterize the proteome of cartilage without in vitro culturing which could obfuscate physiological differences. The definition of unique OA-associated proteins described here provides significant mechanistic insights into OA by corroborating previously suggested mechanisms and by defining unique players with roles yet to be defined in disease pathogenesis. (c) 2008 Elsevier Masson SAS. All rights reserved.