Conclusion P gingivalis is an opportunistic, intracellular patho

Conclusion P. gingivalis is an opportunistic, intracellular pathogen that survives for

extended periods of time within gingival epithelial cells without causing excessive harm to the host and thus provides a window into host cell adaptive responses by pathogens [3–5]. Re-analysis of whole cell proteomics data using the recently published strain specific genome annotation for ATCC 33277 allowed several novel conclusions. As expected, the strain specific annotation yielded better overall proteome coverage and sampling depth at the level of the number of proteins identified. However, most of the overall trends identified for major P. gingivalis virulence factors and other proteins using the W83 genome annotation remain unchanged, showing the viability of employing similar annotations when a strain specific sequence is unavailable. buy BAY 1895344 This observation is especially important for oral and gut microbes, where a rapidly increasing body of genomic and RNA-Seq data suggests that genomic re-arrangements in the absence of major changes in amino acid Selleckchem PF2341066 sequence for the expressed proteins may be

a widespread occurrence. Although some differences in protein primary structure exist among P. gingivalis strains [30], the primary differences observed by Naito et al. are extensive genome re-arrangements [11]. The CX-4945 molecular weight proteomic methods used here are highly sensitive to sequence similarity, but not at all to the order in which genes occur on the chromosome. However, the ways in which proteome data are interpreted in terms of operon and regulon structure are greatly influenced by the physical arrangement Progesterone of the genome. When the data were organized in terms of metabolic pathways the whole cell proteomics analysis revealed what appears to be a nutritionally rich intracellular environment for P. gingivalis. The energy metabolism pathway from the preferred amino acids aspartate/asparagine

showed a significant increase. Transcription and translation proteins also showed significant increases, consistent with energy not being limiting. The production of cytotoxic metabolic byproducts also appears to shift in internalized cells, reducing production of butyrate and increasing production of propionate. This may be simply a byproduct of metabolic shifts, or it may play a role in P. gingivalis adaptive response to internalization. Methods Proteomic methods The bacterial and gingival cell culturing, sample preparation, proteome extraction, proteolytic digestion, HPLC pre-fractionation, 2-D capillary HPLC [31, 32], LTQ linear ion trap mass spectral data acquisition parameters, Sequest database searching [33], DTASelect [34]in silico assembly of the P.

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