BGI-Shenzhen , Beishan Industrial Zone 11th building, Yantian District, Shenzhen, Guangdong 518083, China.
James D. Watson Institute of Genome Sciences, Hangzhou 310008, China.
J Proteome Res. 2017 Dec 1;16(12):4330-4339. doi: 10.1021/acs.jproteome.7b00353. Epub 2017 Oct 26.
Human Proteome Project (HPP) aims at mapping entire human proteins with a systematic effort upon all the emerging techniques, which would enhance understanding of human biology and lay a foundation for development of medical applications. Until now, 2563 missing proteins (MPs, PE2-4) are still undetected even using the most sensitive approach of protein detection. Herein, we propose that enrichment of low-abundance proteins benefits MPs finding. ProteoMiner is an equalizing technique by reducing high-abundance proteins and enriching low-abundance proteins in biological liquids. With triton X-100/TBS buffer extraction, ProteoMiner enrichment, and peptide fractionation, 20 MPs (at least two non-nested unique peptides with more than eight a.a. length) with 60 unique peptides were identified from four human tissues including eight membrane/secreted proteins and five nucleus proteins. Then 15 of them were confirmed with two non-nested unique peptides (≥9 a.a.) identified by matching well with their chemically synthetic peptides in PRM assay. Hence, these results demonstrated ProteoMiner as a powerful means in discovery of MPs.
人类蛋白质组计划（HPP）旨在利用所有新兴技术对整个人类蛋白质进行系统绘制，这将增进人们对人类生物学的理解，并为医疗应用的发展奠定基础。迄今为止，即使使用最敏感的蛋白质检测方法，仍有 2563 种缺失蛋白（MPs，PE2-4）未被检测到。在此，我们提出富集低丰度蛋白质有助于发现 MPs。ProteoMiner 是一种均衡技术，可减少生物液中高丰度蛋白质并富集低丰度蛋白质。通过 Triton X-100/TBS 缓冲液提取、ProteoMiner 富集和肽分级，从包括 8 种膜/分泌蛋白和 5 种核蛋白在内的 4 个人体组织中鉴定出 20 种 MPs（至少有两条非嵌套的独特肽，长度超过 8 个氨基酸），有 60 个独特肽。然后，通过与 PRM 测定中化学合成肽相匹配，用两条非嵌套的独特肽（≥9 个氨基酸）确认了其中的 15 个。因此，这些结果表明 ProteoMiner 是发现 MPs 的有力手段。
