Proteomic analysis of human cervico-vaginal fluid.
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.
Shaw
Julie L V
JL
Department of Pathology and Laboratory Medicine, Mount Sinai Hospital, Toronto, Ontario, Canada.
Smith
Christopher R
CR
Diamandis
Eleftherios P
EP
D015415:Biomarkers / Q000032:analysis; D002448:Cell Adhesion; D002582:Cervix Mucus / Q000737:chemistry* / Q000378:metabolism; D002852:Chromatography, Ion Exchange; D002853:Chromatography, Liquid; D003599:Cytoskeleton / Q000737:chemistry / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D045604:Extracellular Fluid / Q000737:chemistry* / Q000378:metabolism; D005260:Female; D006801:Humans; D013058:Mass Spectrometry; D010447:Peptide Hydrolases / Q000032:analysis; D011506:Proteins / Q000032:analysis; D020543:Proteome / Q000032:analysis*; D040901:Proteomics; D014621:Vagina / Q000737:chemistry*
D015415:Biomarkers; D011506:Proteins; D020543:Proteome; D010447:Peptide Hydrolases