6G) However, this cleavage product of PKC-α was not evident in e

6G). However, this cleavage product of PKC-α was not evident in earlier experiments when macrophages PD0332991 solubility dmso were infected with mycobacteria. We speculated that this could be either due to the lower level/less accumulation of PknG (and degradation product) in macrophages as compared to exogenous

addition or may be the further degradation of cleaved products within the cell. Therefore when the proteins were incubated in higher amounts the cleavage product could be seen. Thus we concluded that the presence of PknG in macrophages either with mycobacteria or as a protein, precisely control PKC-α. Moreover, when pathogenic mycobacteria reside in macrophages it raises the level of PknG [Fig. 4C] which strengthens the understanding that more inactivation of PKC-α may be possible. Hence, this study is first to report that PknG downregulates PKC-α and their association discriminates the fates of pathogenic and nonpathogenic mycobacteria in macrophages. Recently,

L. donovani GP63 has been shown to proteolytically cleave many host proteins resulting in the inactivation of MAPKs [32] suggesting that cleavage of host proteins is a defense strategy utilized by intracellular pathogens. During tuberculosis, host defense may be determined, in part, LY2109761 solubility dmso by the capaCity of macrophages to bind and ingest Mtb. Phagocytosis by macrophages represents the early step of the mycobacterial infection. It is governed both by the nature of the host receptors used and the ligands exposed on the bacteria as well as the environment where host cells encounter mycobacteria [[24, 25], and [33–36]]. Final outcome of the infection is determined by cumulative MK-4827 in vivo effect of all these factors. Conclusions Expression of PknG in BCG, Ra and Rv but not in MS represents an interesting example of evolution

where pathogen has developed strategies for modulation of host molecule to avoid uptake and killing by the entities designed for their killing. Interestingly, PknG directs the downregulation of PKCα and further negotiating the uptake and survival of Amoxicillin mycobacteria. Our data clearly and for the first time reveal that pathogenic mycobacteria downregulates PKCα predominantly to avoid phagocytosis and killing by macrophages. Detailed understanding of the events leading to the downregulation of PKCα by PknG inside host cells open a new chapter which may further project to the identification of new therapeutic targets for mycobacterial infections. Methods Reagents Antibodies against PKCs and phospho-PKCs were purchased through Santa Cruz Biotech Inc. and Cell Signaling Technologies, USA, respectively. Horseradish peroxidase-linked secondary antibodies, polyvinylidene difluoride membrane, RPMI-1640, DMEM, HEPES, sodium bicarbonate, Imidazole, IPTG and Protein G Sepharose beads were purchased from Sigma chemicals. Enhanced chemiluminescence kit (ECL) was from GE Healthcare.

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