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优化人类关节蛋白质组二维凝胶电泳分析的策略。

Strategies to optimize two-dimensional gel electrophoresis analysis of the human joint proteome.

机构信息

Proteomics Unit and Associated Node to ProteoRed, Osteoarticular and Aging Research Lab, INIBIC-Hospital Universitario A Coruña, A Coruña, Spain.

出版信息

Talanta. 2010 Feb 15;80(4):1552-60. doi: 10.1016/j.talanta.2009.05.022. Epub 2009 May 22.

Abstract

Due to the complex structure of the articular joint, it requires great effort to fully understand joint disease pathogenesis. The proteomic analysis of articular joint tissues could contribute greatly to our insight into the endogenous control mechanisms of matrix turnover and the unravelling of the molecular and cellular mechanisms involved in the progression of the arthritides. To date, most proteome analysis strategies use the two-dimensional gel electrophoresis (2-DE) technique to separate proteins according to their isoelectric point, molecular mass, solubility and relative abundance. In this work, we describe optimization of human joint sample preparation techniques to obtain high quality 2-DE maps of human joint tissues (cartilage and synovium), cells (chondrocytes and synoviocytes) and synovial fluid. These techniques improve the performance of gel-based differential proteomic analysis, and facilitate the application of proteomics to rheumatology studies.

摘要

由于关节的复杂结构,要全面了解关节疾病的发病机制需要付出巨大的努力。关节组织的蛋白质组学分析可以极大地帮助我们深入了解基质周转的内源性控制机制,并揭示关节炎进展中涉及的分子和细胞机制。迄今为止,大多数蛋白质组分析策略都使用二维凝胶电泳 (2-DE) 技术根据等电点、分子量、溶解度和相对丰度来分离蛋白质。在这项工作中,我们描述了优化人关节样品制备技术,以获得高质量的人关节组织(软骨和滑膜)、细胞(软骨细胞和滑膜细胞)和滑液的 2-DE 图谱。这些技术提高了基于凝胶的差异蛋白质组分析的性能,并促进了蛋白质组学在风湿病研究中的应用。

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