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首页> 美国卫生研究院文献>Scientific Reports >Evaluating the potential of residual Pap test fluid as a resource for the metaproteomic analysis of the cervical-vaginal microbiome
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Evaluating the potential of residual Pap test fluid as a resource for the metaproteomic analysis of the cervical-vaginal microbiome

机译：评估残留子宫颈抹片检查液作为宫颈阴道微生物组的蛋白质组学分析资源的潜力

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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.
机译：人类的子宫颈阴道区域含有源自微生物的蛋白质，这些蛋白质可能会阻止妇女或使妇女患上妇科病。液体巴氏试验固定剂是用于分析微生物群落和微生物-宿主相互作用的未开发资源。以前，我们表明健康女性丢弃的Pap测试中残留的​​无细胞固定剂可用于基于质谱（MS）的宫颈阴道蛋白的蛋白质组学鉴定。在这项研究中，我们重新处理了这些MS原始数据文件，以进行元蛋白质组学分析，以表征子宫颈阴道区域的微生物群落组成和微生物蛋白的功能。这是通过开发定制的蛋白质序列数据库实现的，该数据库包含可能存在于阴道中的微生物。在银河蛋白质组学平台上使用两步搜索方法，针对蛋白质FASTA数据库搜索了高质量数据。通过MEGAN6（MetaGenomeAnalyzer）分析数据，以进行系统发育和功能表征。我们从各种细菌门和念珠菌中鉴定了300多种独特的肽。鉴定了与碳水化合物代谢，氧化还原和转运相关的蛋白质对应的肽。通过在Pap测试上清液中鉴定微生物肽，可能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。

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