It is clear that the cytosolically expressed Inc proteins from C. pneumoniae species failed to significantly alter the susceptibility of the transfected cells to the subse quent C. pneumoniae infection. Discussion In the current study, we have provided the first experimen tal evidence demonstrating that the hypothetical selleckchem Enzastaurin proteins Cpn0146 147 are Inc proteins while Cpn0284 and 0285 are associated with RBs inside the inclusions although all 4 proteins were predicted to be in the inclusion mem brane based on their N terminal bi lobed hydrophobic Inhibitors,Modulators,Libraries motifs. Although the antibodies used for localiz ing these four proteins were raised with fusion proteins, we have convincingly demonstrated that the anti fusion protein antibodies specifically recognized the correspond ing endogenous antigens during C.
pneumoniae infection. It is interesting that Cpn0147 was detected as early as 6 hours while IncA was only detected 12 hours and Cpn0146, 0284 0285 24 hours after infection. These differential protein expression patterns suggest that Cpn0147 and IncA are early genes while Cpn0146, 0284 0285 late genes although further monitoring transcripts is required for providing Inhibitors,Modulators,Libraries evidence to support the conclu sion. Although Cpn0146 and 0147 are encoded in the same gene cluster and oriented in the same direction in the C. pneumoniae genome, Cpn0147 is expressed much earlier than Cpn0146, suggesting that the expression of these two genes are regulated independently. Regardless of the difference in the timing of expression, both Cpn0146 and 0147 proteins remained in the inclusion membrane once detected.
However, the RB Inhibitors,Modulators,Libraries proteins Cpn0284 and 0285 fluctuated along the infection cycle with strong signals in small inclusions and no or very low signals in large inclusions between 24 and 96 hours Inhibitors,Modulators,Libraries and reappeared in large inclusions by 120 hours after infec tion, suggesting that Cpn0284 0285 are RB specific pro teins although more biochemical assays are required for confirming this conclusion. Since the identification of the first chlamydial inclusion membrane protein, various features have been assigned to chlamydial inclusion membrane proteins, including localization in the inclusion membrane, acces sibility to host immunoproteasomal processing, secreta bility Inhibitors,Modulators,Libraries by various heterologous type III secretion systems, bi lobed hydrophobic motifs, ER like distribution and the ability to render host cell resistance to subsequent chlamydial infection when pre expressed in host cell cytosol.
Here, we systematically analyzed four predicted Gemcitabine clinical Inc proteins encoded by two separate gene clusters in the C. pneumoniae genome. We found that Cpn0146 0147 but not Cpn0284 0285 were localized in the inclusion membrane during C. pneumoniae infection and co local ized with ER when expressed via transgenes, which sug gests that the ability of C.