Global Survey of the Bovine Salivary Proteome: Integrating Multidimensional Prefractionation, Targeted, and Glycocapture Strategies
Global Survey of the Bovine Salivary Proteome: Integrating Multidimensional Prefractionation, Targeted, and Glycocapture Strategies
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DOI：
10.1021/pr200516d
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发表时间：
2011-11-01
影响因子：
4.4
通讯作者：
McDonagh, Matthew B.
中科院分区：
文献类型：
--
作者：
Ang, Ching-Seng;Binos, Steve;McDonagh, Matthew B.
唾液很容易以非侵入性的方式从大量动物中获得，并且含有多种多样的化合物，包括激素、代谢物和蛋白质，这些化合物可能是健康和疾病生物标志物的良好来源。在这里，我们使用多维预分级、靶向和糖捕获方法的组合来分析牛唾液蛋白质组。非靶向方法使用四种不同的分离方法，包括SDS-PAGE、离胶分级分离、RP-HPLC和SCX-HPLC。在靶向方法中，我们采用了基于假设的方法，仅从计算机数据中选择细胞外蛋白。最后，酰肼捕获方法不仅使我们能够识别以前的N-连接的糖蛋白，但它也提供了一个选择性的富集过程，用于低丰度蛋白质的识别。总之，这三种不同的方法鉴定了402种唾液蛋白和45种N-连接糖蛋白。这些蛋白质中的大量先前在牛唾液中未被表征。迄今为止，这是全球最大的牛唾液蛋白质组调查，并扩大了这种液体的诊断效用的潜力，以指导寻找健康特征生物标志物的实验的发展（即，抗病性）以及奶牛和肉牛的饲料转化效率和生产力性状。
Saliva is easily obtainable from a large number of animals in a non-invasive manner and contains a wide diversity of compounds including hormones, metabolites, and proteins that may be a good source of biomarkers of health and disease. Here we have used a combination of multidimensional prefractionation, targeted, and glycocapture methodologies to profile the bovine salivary proteome. The nontargeted approach used four different separation methodologies consisting of SDS-PAGE, Off-gel fractionation, RP-HPLC, and SCX-HPLC. In the targeted approach, we've employed a hypothesis-based methodology by only selecting extracellular proteins from in silico data. Finally, the hydrazide capture methodology not only enabled us to identify formerly N-linked glycoproteins but it also provided a selective enrichment process for the identification of low abundance proteins. Together, the three different approaches identified 402 salivary proteins and 45 N-linked glycoproteins. A large number of these proteins have previously been uncharacterized in bovine saliva. To date, this is the largest global survey of the bovine salivary proteome and expands the potential of the diagnostic utility of this fluid to guide development of experiments seeking biomarkers for health traits (i.e., disease resistance) as well as feed conversion efficiency and productivity traits in dairy and beef cattle.