Mass Spectrometry Identification of Outer Membrane Proteins of Prevotella intermedia 17, Interacting with Host Cells. // Acta Scientific Cancer Biology, 2018, V. 2 (3), PР. 02-13статья
Дата последнего поиска статьи во внешних источниках: 1 июня 2022 г.
Аннотация:Bacterial pathogens attach to the cell surface of host and then induce host cells or bacterial changes to gain entry to the host cells. An analysis of bacteria present in the mouth showed that some types of bacteria that lead to periodontal disease were associated with higher risk of esophageal cancer. Adhesin bacteria-host proteins interactions are necessary for invasion and bacterial colonization of host tissues. Adherence of P.intermedia 17 to the host cells is the initial and crucial step in development of periodontal disease. Identification and characterization of outer membrane proteins (Omps) P.intremedia 17 associated with cell surface host proteins is important step in investigation of molecular mechanism of host-bacteria interactions.
Here we used sensitive fluorescent labeling Omps of P.intermedia 17, followed incubation with host cells (oral epithelial and human fibroblasts), 1D gel separations and mass spectrometric identification (LC-MS/MS) of cell surface proteins. We identified 7 Omps of P.intermedia 17, interacting with oral epithelial cells. Two from them, Omp 41(PIN_A1455, immunoreactive 43 kDa antigen PG32, putative) and major Omp (PIN_A0102, immunoreactive 42 kDa antigen PG33, putative) were determined with high sequence coverage and top score of peptides match. Effect of heat on host-bacteria interaction has been studied. Similar native and denaturated Omps, bound with host cells, were identified by LC-MS/MS analysis. Denaturated Omps demonstrated the lower binding affinity in comparison with native. Nine Omps were detected, associated with human fibroblasts. Cell surface Omps bound to human fibroblasts were determined, using “shaving” approach. Hypothetical protein (PIN_A0324) has been identified as protein, interacting with cell surface proteins from human fibroblasts.
