Proteomic analysis of human cervico-vaginal fluid
Proteomic analysis of human cervico-vaginal fluid
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DOI：
10.1021/pr0701658
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发表时间：
2007-01-01
影响因子：
4.4
通讯作者：
Diamandis, Eleftherios. P.
中科院分区：
文献类型：
--
作者：
Shaw, Julie L. V.;Smith, Christopher R.;Diamandis, Eleftherios. P.
人宫颈阴道液（CVF）是源自阴道、子宫颈、子宫内膜和输卵管的液体的混合物。CVF已被证明在保护阴道免受感染方面发挥重要作用。我们使用“自下而上”的蛋白质组学方法来表征人类CVF的蛋白质库。我们采用了一维sds - page （1D-SDS-PAGE）和强阳离子交换色谱两种不同的样品前分离方法，然后采用LC-MS/MS和生物信息学分析。我们一共鉴定出685种蛋白质。与1D-SDS-PAGE相比，强阳离子交换色谱预分离可以鉴定出更多的蛋白质。根据基因组本体（GO）分类，细胞外或膜蛋白占鉴定蛋白质的30%。我们证实了防御相关蛋白的存在，如触珠蛋白、防御蛋白和乳铁蛋白；并鉴定出一些新的药物，如灭虫素和杀皮素。我们还发现了许多丝氨酸和半胱氨酸蛋白酶，包括kallikrein家族的6个成员（KLKs 6、7、10、11、12和13）。同样的KLKs也被ELISA定量地证实。CVF蛋白质组的知识将有助于发现妇科恶性肿瘤和感染的潜在生物标志物，并为其生理功能提供额外的线索。
Human cervico-vaginal fluid (CVF) is a mixture of fluids originating from the vagina, cervix, endometrium, and oviduct. CVF has been shown to play an important role in protecting the vagina from infection. We used "bottom-up" proteomic approaches to characterize the protein repertoire of human CVF. We applied two different sample prefractionation methods, one-dimensional-SDS-PAGE (1D-SDS-PAGE) and strong cation-exchange chromatography, followed by LC-MS/MS and bioinformatic analysis. We identified a total of 685 proteins. Strong cation-exchange chromatography prefractionation resulted in a larger number of proteins identified when compared with 1D-SDS-PAGE. Extracellular or membrane proteins made up 30% of the proteins identified, according to Genome Ontology (GO) classifications. We confirmed the presence of defense-related proteins, such as haptoglobin, defensins, and lactoferrin; and identified new ones such as azurocidin and dermcidin. We also identified many serine and cysteine proteases, including 6 members of the kallikrein family (KLKs 6, 7, 10, 11, 12, and 13). The same KLKs were also confirmed quantitatively by ELISA assays. Knowledge of the CVF proteome will aid in the discovery of potential biomarkers for gynecological malignancies and infections and provide additional clues for its physiological functions.