Zhou Lei, Beuerman Roger W, Foo Yonghwee, Liu Shouping, Ang Leonard P K, Tan Donald T H
Singapore Eye Research Institute, Singapore.
Ann Acad Med Singap. 2006 Jun;35(6):400-7.
The proteins found in tears play an important role in maintaining the ocular surface and changes in tear protein components may reflect changes in the health of the ocular surface. Proteomics provides a comprehensive approach for cataloguing all the proteins of the tear proteome, which will help to elucidate disease pathogenesis, make clinical diagnoses and evaluate the influence of medications on the structure, composition and secretion of tear proteins. In this study, an alternative proteomic strategy was investigated to explore the human tear proteome.
Tear samples were obtained from patients who had pterygium and were collected on the first day and third day after pterygium surgery. Tears pooled from 6 patients were used in the analysis. Reverse-phase high-pressure liquid chromatograph (RPHPLC) was used as the first step to separate intact proteins into 21 peaks. Each fraction was then tryptic-digested and analysed by nanoLC-nano-ESI-MS/MS to characterise the protein components in each fraction.
In total, 60 tear proteins were identified with high confidence, including well-known abundant tear proteins, and tear-specific proteins such as lacritin and proline-rich proteins. Among them, proline-rich protein 5 was found for the first time in tear fluid. A large number of plasma proteins were also observed in tear fluid.
The results showed that the proteomic strategy used in this study was successfully applied to analyse tear proteome.
泪液中的蛋白质在维持眼表健康方面发挥着重要作用，泪液蛋白质成分的变化可能反映眼表健康状况的改变。蛋白质组学为编目泪液蛋白质组中的所有蛋白质提供了一种全面的方法，这将有助于阐明疾病发病机制、进行临床诊断以及评估药物对泪液蛋白质的结构、组成和分泌的影响。在本研究中，研究了一种替代蛋白质组学策略来探索人类泪液蛋白质组。
从患有翼状胬肉的患者中获取泪液样本，并在翼状胬肉手术后第一天和第三天收集。将6名患者的泪液混合用于分析。第一步使用反相高压液相色谱（RPHPLC）将完整蛋白质分离成21个峰。然后对每个馏分进行胰蛋白酶消化，并通过纳升液相色谱-纳升电喷雾串联质谱（nanoLC-nano-ESI-MS/MS）进行分析，以表征每个馏分中的蛋白质成分。
总共高可信度地鉴定出60种泪液蛋白质，包括众所周知的丰富泪液蛋白质以及泪液特异性蛋白质，如乳铁蛋白和富含脯氨酸的蛋白质。其中，富含脯氨酸的蛋白质5首次在泪液中被发现。在泪液中还观察到大量血浆蛋白质。
结果表明本研究中使用的蛋白质组学策略成功应用于分析泪液蛋白质组。
