Sequential Precipitation and Delipidation Enables Efficient Enrichment of Low-Molecular Weight Proteins and Peptides from Human Plasma.,Journal of Proteome Research

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Sequential Precipitation and Delipidation Enables Efficient Enrichment of Low-Molecular Weight Proteins and Peptides from Human Plasma.
Journal of Proteome Research ( IF 3.8 ) Pub Date : 2020-07-03 , DOI: 10.1021/acs.jproteome.0c00232
Na Li _{1,

2} , Yue Zhou _{1,

3} , Jifeng Wang ₁ , Lili Niu ₁ , Qing Zhang _{1,

2} , Lang Sun _{1,

2} , Xiang Ding ₁ , Xiaojing Guo _{1,

2} , Zhensheng Xie ₁ , Nali Zhu ₁ , Mengmeng Zhang ₁ , Xiulan Chen _{1,

2} , Tanxi Cai _{1,

2} , Fuquan Yang _{1,

2}

Affiliation

Low-molecular weight proteins and peptides (LMWPs, <30 kDa) in human plasma serve as potential biomarkers or drug targets and are endowed with desirable traits for biological and clinical studies. However, the identification of LMWPs from plasma is retarded by high-abundance proteins, high-molecular weight proteins, and lipids. Here, we present a sequential precipitation and delipidation (SPD) method for the efficient enrichment of LMWPs based on methyl-tert-butyl ether/methanol/water systems. The enriched LMWP sample was analyzed by single-shot liquid chromatography–tandem mass spectrometry employing both HCD and EThcD without tryptic digestion, and 725 peptides were identified on average. The LMWP sample was also digested and analyzed using a bottom-up proteomics pipeline, and 289 proteins were identified, of which 129 (44.6%) proteins were less than 30 kDa and lipoprotein-associated proteins were significantly enriched. Additionally, 25 neuropeptides and 19 long noncoding RNA-encoded polypeptides were identified. Taken together, the SPD method shows good sensitivity and reproducibility when compared with other enrichment methods and has great potential for clinical biomarker discovery and application.

中文翻译：

顺序沉淀和脱脂使人血浆中的低分子量蛋白质和肽有效富集。

人血浆中的低分子量蛋白质和肽（LMWPs，<30 kDa）可作为潜在的生物标记物或药物靶标，并具有生物学和临床研究所需的特征。但是，血浆中LMWP的鉴定受到高丰度蛋白质，高分子量蛋白质和脂质的阻碍。在这里，我们提出了一种顺序沉淀和脱脂（SPD）方法，用于基于甲基叔丁基醚的LMWP有效富集-丁基醚/甲醇/水系统。富集的LMWP样品通过使用HCD和EThcD的单次液相色谱-串联质谱分析而没有胰蛋白酶消化，进行了分析，平均鉴定出725种肽。还使用自下而上的蛋白质组学管线对LMWP样品进行了消化和分析，鉴定出289种蛋白质，其中129种（44.6％）蛋白质小于30 kDa，脂蛋白相关的蛋白质显着富集。另外，鉴定出25种神经肽和19种长的非编码RNA编码的多肽。综上所述，与其他富集方法相比，SPD方法显示出良好的灵敏度和可重复性，在临床生物标志物的发现和应用方面具有巨大的潜力。

更新日期：2020-08-08

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