Images were captured at magnification of 6000x (C) Localization by immunoblotting
Images were captured at magnification of 6000x (C) Localization by immunoblotting. was initially investigated by analyzing the presence of SPFH2a, oligomerization domains in the concerned gene and NfeD domain name in the adjacent upstream gene. After investigating 300 PB, four were found to harbor RMP. Among them, domains of Bas0525 (FlotP) of (as the bioterror 3-O-(2-Aminoethyl)-25-hydroxyvitamin D3 agent, it was selected as a model for further characterization of rafts in PB. and analysis showed significant similarity of FlotP with numerous attributes of Flotillin-1. Its punctate distribution on membrane with exclusive localization in detergent resistant membrane fraction; strongly favors presence of raft with RMP FlotP in BA. Furthermore, significant effect of Zaragozic acid (ZA), a raft associated lipid biosynthesis inhibitor, on several patho-physiological attributes of BA such as growth, morphology, membrane rigidity etc., were also observed. Specifically, a considerable decrease in membrane rigidity, strongly recommended presence of an unknown raft associated lipid molecule on membrane of BA. In addition, treatment with ZA decreased secretion of anthrax toxins and FlotP expression, suggesting potential role of raft in pathogenesis and physiology of BA. Thus, the present study not only suggest 3-O-(2-Aminoethyl)-25-hydroxyvitamin D3 the presence and role of raft like entity in pathophysiology of BA but also its possible use for the development of novel drugs or vaccines against anthrax. (Zhang et al., 2005; Meile et al., 2006; Donovan and Bramkamp, 2009; Lpez and Kolter, 2010). The role of these bacterial rafts in membrane heterogeneity, sporulation, biofilm formation etc., (Donovan and Bramkamp, 2009; Bramkamp and Lopez, 2015) raises 3-O-(2-Aminoethyl)-25-hydroxyvitamin D3 the idea that these might be the niche for numerous proteins involved in crucial bacterial physiological processes. Moreover, in bacteria, lipid raft disruption interferes with vital cellular processes without affecting cell viability and thus, is less likely to exhibit spontaneous mutation-acquired resistance against anti-raft compounds like statins that make these drugs less antimicrobial resistant (Donovan and Bramkamp, 2009; Bramkamp and Lopez, 2015). Commercially available anti-raft compounds have already been proved to be nontoxic to humans even at higher concentrations (Bramkamp and Lopez, 2015). In addition, rafts are also known to harbor sterol like molecules. Therefore, inhibitors of enzymes involved in rafts associated lipid biosynthesis have also been shown to affect microdomain organization and thus, physiology of the organism (Lpez and Kolter, 2010). Hence, investigating presence of rafts in pathogenic microbes is likely to reveal hidden insights into bacterial pathophysiology to Rabbit polyclonal to FASTK develop novel drugs or anti-raft compounds that may possess broad spectrum activity, less antimicrobial resistance and minimal host side effects. Although, variations might exist in microdomains associated proteins or lipids, but features of Flotillin-like proteins remain conserved throughout. Thus, characterization of Flotillin appears to be the most affordable approach to explore microdomains in bacteria. In this study, we performed analysis to find protein(s) in bacterial pathogens with various primary and secondary structural attributes of Flotillin. The high identity of BAS0525 (FlotP) of to above attributes of Flotillin-1 and importance of anthrax in public health globally prompted us to characterize FlotP (Simons and Sampaio, 2011). Also, it has been seen that biosynthesis of bacterial raft associated lipids involve squalene synthase (Lpez and Kolter, 2010) whose known inhibitor Zaragozic acid (ZA) was found to impede the presence and 3-O-(2-Aminoethyl)-25-hydroxyvitamin D3 formation of bacterial raft. Thus, to claim presence of raft, a parallel investigation of raft associated lipid molecule in bacteria would be highly recommended. Therefore, in order to further explore raft, effect of this inhibitor on several pathophysiological attributes of were also studied. Materials and methods identification of flotillin homolog and its related gene in pathogens For the identification and selection of the most significant Flotillin homolog harboring pathogens from around 300 different pathogenic bacterial strains, following steps were followed. All sequences were retrieved from KEGG database. Initially, orthologs of SPFH2a domain name made up of homolog of prokaryotic Flotillin were searched by Clustal W.