Biosciences Research Division, Dairy Futures Cooperative Research Centre, Bundoora, Victoria, Australia.
J Proteome Res. 2011 Nov 4;10(11):5059-69. doi: 10.1021/pr200516d. Epub 2011 Sep 28.
Saliva is easily obtainable from a large number of animals in a noninvasive 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.
唾液可以通过非侵入性的方式从大量动物中获得，其中含有广泛多样的化合物，包括激素、代谢物和蛋白质，这些可能是健康和疾病生物标志物的良好来源。在这里，我们结合了多维预分级、靶向和糖捕获方法来分析牛唾液蛋白质组。非靶向方法使用了四种不同的分离方法，包括 SDS-PAGE、Off-gel 分级、RP-HPLC 和 SCX-HPLC。在靶向方法中，我们仅根据计算机数据选择细胞外蛋白质，采用了基于假设的方法。最后，酰肼捕获方法不仅使我们能够鉴定以前的 N 连接糖蛋白，而且还提供了一种用于鉴定低丰度蛋白质的选择性富集过程。这三种不同的方法共鉴定出 402 种唾液蛋白和 45 种 N 连接糖蛋白。这些蛋白质中有很大一部分以前在牛唾液中没有被描述过。迄今为止，这是对牛唾液蛋白质组的最大规模的全球调查，扩大了这种液体的诊断效用的潜力，可用于指导开发用于寻找健康特征（例如疾病抗性）以及奶牛和肉牛的饲料转化率效率和生产力特征的生物标志物的实验。
