The human cervical-vaginal area contains proteins derived from microorganisms that may prevent or predispose women to gynecological conditions. The liquid Pap test fixative is an unexplored resource for analysis of microbial communities and the microbe-host interaction. Previously, we showed that the residual cell-free fixative from discarded Pap tests of healthy women could be used for mass spectrometry (MS) based proteomic identification of cervical-vaginal proteins. In this study, we reprocessed these MS raw data files for metaproteomic analysis to characterize the microbial community composition and function of microbial proteins in the cervical-vaginal region. This was accomplished by developing a customized protein sequence database encompassing microbes likely present in the vagina. High-mass accuracy data were searched against the protein FASTA database using a two-step search method within the Galaxy for proteomics platform. Data was analyzed by MEGAN6 (MetaGenomeAnalyzer) for phylogenetic and functional characterization. We identified over 300 unique peptides from a variety of bacterial phyla and Candida. Peptides corresponding to proteins involved in carbohydrate metabolism, oxidation-reduction, and transport were identified. By identifying microbial peptides in Pap test supernatants it may be possible to acquire a functional signature of these microbes, as well as detect specific proteins associated with cervical health and disease.

中文翻译：

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评估残留巴氏试验液作为宫颈阴道微生物组蛋白质组分析资源的潜力。


人类宫颈阴道区域含有源自微生物的蛋白质，可以预防或使女性易患妇科疾病。液体巴氏试验固定剂是一种未经开发的资源，用于分析微生物群落和微生物-宿主相互作用。此前，我们表明，健康女性废弃巴氏试验中残留的无细胞固定剂可用于基于质谱（MS）的宫颈阴道蛋白质的蛋白质组学鉴定。在这项研究中，我们重新处理这些 MS 原始数据文件进行宏蛋白质组学分析，以表征宫颈阴道区域微生物群落的组成和微生物蛋白质的功能。这是通过开发包含阴道中可能存在的微生物的定制蛋白质序列数据库来实现的。使用 Galaxy 蛋白质组学平台内的两步搜索方法在蛋白质 FASTA 数据库中搜索高质量准确数据。通过 MEGAN6（MetaGenomeAnalyzer）分析数据以进行系统发育和功能表征。我们从多种细菌门和念珠菌中鉴定出 300 多种独特的肽。鉴定出与参与碳水化合物代谢、氧化还原和运输的蛋白质相对应的肽。通过鉴定巴氏试验上清液中的微生物肽，可以获得这些微生物的功能特征，并检测与宫颈健康和疾病相关的特定蛋白质。