Stastna Miroslava, Behrens Ashley, Noguera Guillermo, Herretes Samantha, McDonnell Peter, Van Eyk Jennifer E
Department of Medicine, Johns Hopkins Bayview Proteomics Center, Johns Hopkins University, Baltimore, MD 21224, USA.
Proteomics. 2007 Dec;7(23):4358-75. doi: 10.1002/pmic.200700300.
There are several physiological roles postulated for aqueous humor, a liquid located in the anterior and posterior chamber of the eye, such as maintenance of the intraocular pressure, provision of nutrients, and removal of metabolic waste from neighboring tissues and provision of an immune response and protection during inflammation and infection. To link these function to specific or classes of proteins, identification of the aqueous humor proteome is essential. Aqueous humor obtained from healthy New Zealand white rabbits was analyzed using three synergistic protein separation methods: 1-D gel electrophoresis, 2-DE, and 1-DLC (RPLC) prior to protein identification by MS. As each of these separation methods separates intact proteins based on different physical properties (pIs, molecular weights, hydrophobicity, solubility, etc.) the proteome coverage is expanded. This was confirmed, since overlap between all three separation technologies was only about 8.2% with many proteins found uniquely by a single method. Although the most dominant protein presented in normal aqueous humor is albumin, by using this extensive separation/MS strategy, additional proteins were identified in total amount of 98 nonredundant proteins (plus an additional ten proteins for consideration). This expands the current protein identifications by approximately 65%. The aqueous humor proteome comprises a specific selection of cellular and plasma based proteins and can almost exclusively be divided into four functional groups: cell-cell interactions/wound healing, proteases and protease inhibitors, antioxidant protection, and antibacterial/anti-inflammatory proteins.
房水是一种位于眼球前房和后房的液体，人们推测它具有多种生理作用，比如维持眼压、提供营养、清除邻近组织的代谢废物，以及在炎症和感染期间提供免疫反应和保护。为了将这些功能与特定的蛋白质或蛋白质类别联系起来，鉴定房水蛋白质组至关重要。在通过质谱进行蛋白质鉴定之前，使用三种协同的蛋白质分离方法（一维凝胶电泳、二维电泳和一维液相色谱（反相液相色谱））对从健康新西兰白兔获得的房水进行了分析。由于这些分离方法中的每一种都是基于不同的物理性质（等电点、分子量、疏水性、溶解度等）来分离完整蛋白质的，因此蛋白质组覆盖范围得以扩大。这一点得到了证实，因为所有三种分离技术之间的重叠仅约为8.2%，许多蛋白质是通过单一方法独特发现的。虽然正常房水中最主要的蛋白质是白蛋白，但通过这种广泛的分离/质谱策略，总共鉴定出了98种非冗余蛋白质（另外还有十种蛋白质供考虑）。这使目前的蛋白质鉴定数量增加了约65%。房水蛋白质组包括特定选择的基于细胞和血浆的蛋白质，几乎可以完全分为四个功能组：细胞间相互作用/伤口愈合、蛋白酶和蛋白酶抑制剂、抗氧化保护以及抗菌/抗炎蛋白。
