Hebei Province Key Lab of Research and Application on Microbial Diversity, College of Life Sciences , Hebei University , Baoding , Hebei 071002 , China.
State Key Laboratory of Proteomics, Beijing Proteome Research Center, National Center for Protein Sciences (Beijing) , Beijing Institute of Lifeomics , Beijing 102206 , China.
J Proteome Res. 2018 Dec 7;17(12):4171-4177. doi: 10.1021/acs.jproteome.8b00397. Epub 2018 Oct 16.
Subsequent to conducting the Chromosome-Centric Human Proteome Project, we have focused on human testis-enriched missing proteins (MPs) since 2015. For protein coverage to be enhanced, a multiprotease strategy was used for separation of samples by 10% SDS-PAGE. For the separating efficiency to be improved, a high-pH reverse phase (RP) separation strategy was applied to fractionate complex samples in this study. A total of 11,558 proteins was identified, which is the largest proteome data set for single human tissue sample so far. On the basis of this large-scale data set, we verified 14 MPs (PE2) in neXtProt (2018-01) after spectrum quality analysis, isobaric post-translational modification, and single amino acid variant filtering, and synthesized peptide matching. Tissue expression analysis showed that 3 of 14 MPs were testis-specific proteins. Functional analysis showed that 10 of 14 MPs were closely related to liver tumor, liver carcinoma, and hepatocellular carcinoma. Another 100 MPs were listed as candidates but required additional verification information. All MS data sets have been deposited into the ProteomeXchange with the identifier PXD009737.
在开展染色体为中心的人类蛋白质组计划之后，自 2015 年以来，我们一直专注于人类睾丸丰富的缺失蛋白（MPs）。为了提高蛋白质的覆盖率，我们采用了多蛋白酶策略，通过 10%SDS-PAGE 对样品进行分离。为了提高分离效率，本研究应用了高 pH 值反相（RP）分离策略来分离复杂的样品。共鉴定出 11558 种蛋白质，这是迄今为止单个人类组织样本中最大的蛋白质组数据集。基于这个大规模数据集，我们通过谱质量分析、等电点翻译后修饰和单氨基酸变异过滤以及合成肽匹配，在 neXtProt（2018-01）中验证了 14 个 MPs（PE2）。组织表达分析表明，14 个 MPs 中有 3 个是睾丸特异性蛋白。功能分析表明，10 个 MPs 与肝肿瘤、肝癌和肝细胞癌密切相关。还有 100 个 MPs 被列为候选者，但需要额外的验证信息。所有 MS 数据集都已存入 ProteomeXchange，标识符为 PXD009737。
