Li Nan, Wang Nan, Zheng Jing, Liu X Michael, Lever O William, Erickson Paul M, Li Liang
Department of Chemistry, University of Alberta, Edmonton, Alberta, Canada.
J Proteome Res. 2005 Nov-Dec;4(6):2052-61. doi: 10.1021/pr0501970.
Tear proteome profiling may generate useful information for the understanding of the interaction between an eye and its contacting objects, such as a contact lens or a lens implant. This is important for designing improved eye-care devices and maintaining the health of an eye. Proteome profiles of tear fluids may also be used for disease diagnosis and prognosis. However, only a small volume of tear fluid (<5 microL) can be collected in a clinical laboratory under normal operational conditions, which makes proteome profiling a challenge. In this work we apply several proteomic analysis techniques, including gel-based and solution-based approaches with LC-ESI and LC-MALDI MS and MS/MS to gauge the relative merits of producing proteome profiles and to generate as broad a coverage of the tear proteome as possible from this small amount of sample. It is shown that a total of 54 proteins can be confidently identified using less than 5 microL of tear fluid. Of these, 44 proteins can be detected by LC-MALDI MS alone with a consumption of 2 microL of tear fluid. Furthermore, LC-MALDI can be used to determine post-translational modifications (PTMs), such as glycosylation and phosphorylation, without any sample enrichment or treatment. This work represents one of the most extensive proteome profiles (i.e., proteins identified and PTMs characterized) generated from tear fluids using clinically relevant amounts of sample.
泪液蛋白质组分析可为理解眼睛与其接触物体（如隐形眼镜或人工晶状体）之间的相互作用提供有用信息。这对于设计改进的眼部护理设备和维持眼睛健康至关重要。泪液的蛋白质组图谱也可用于疾病诊断和预后评估。然而，在正常操作条件下，临床实验室只能收集少量（<5微升）泪液，这使得蛋白质组分析成为一项挑战。在这项工作中，我们应用了几种蛋白质组分析技术，包括基于凝胶和基于溶液的方法，结合液相色谱-电喷雾电离质谱（LC-ESI）和液相色谱-基质辅助激光解吸电离质谱（LC-MALDI MS）以及串联质谱（MS/MS），以评估生成蛋白质组图谱的相对优势，并从这少量样本中尽可能广泛地覆盖泪液蛋白质组。结果表明，使用不到5微升泪液可以可靠地鉴定出总共54种蛋白质。其中，仅通过LC-MALDI MS就能检测到44种蛋白质，且消耗的泪液仅为2微升。此外，LC-MALDI可用于确定翻译后修饰（PTM），如糖基化和磷酸化，而无需任何样本富集或处理。这项工作代表了使用临床相关量的样本从泪液中生成的最广泛的蛋白质组图谱之一（即鉴定出的蛋白质和表征的PTM）。
