PubMedCrossRef Authors’ buy Veliparib contributions HY designed the experiments and wrote this manuscript; LL performed all phage related experiments; SL analyzed the clinical bacteria strains; HY and SJ supervised the work. The final work was read and accepted by all co-authors.”
“Background Tuberculosis is an airborne infection caused by Mycobacterium tuberculosis. It is estimated that one-third of the world’s population

is latently infected with M. tuberculosis, and that each year about three million people die of this disease. The emergence of drug-resistant stains is further escalating the threat to public health (WHO, 2003). In spite of global research efforts, mechanisms underlying pathogenesis, virulence and persistence of M. tuberculosis infection remain poorly understood [1]. M. tuberculosis is a facultative intracellular pathogen that resides within the host macrophages [2–4]. click here When M. tuberculosis invades host cells, the interface between the Anlotinib manufacturer host and the pathogen includes membrane- and surface proteins likely to be involved in intracellular multiplication and the bacterial response to host microbicidal processes [4]. Recently, the cell wall of M. tuberculosis was reported to posses a true

outer membrane adding more complexity with regard to bacterial-host interactions and also important information relevant for susceptibility to anti-mycobacterial therapies [5–7]. Revealing the composition of the membrane proteome will have an impact on the design and interpretation of experiments aimed at elucidating the translocation Ureohydrolase pathways for nutrients, lipids, proteins, and anti-mycobacterial drugs across the cell envelope. According to bioinformatic predictions, 597 genes (~15%) of the M. tuberculosis H37Rv genome [8, 9], could encode proteins having between 1 and 18 transmembrane α-helical domains (TMH), which interact with the hydrophobic

core of the lipid bilayer. The confirmation of the expression of these genes at the protein level may lead to new therapeutic targets, new vaccine candidates and better serodiagnostic methods. Membrane proteins resolve poorly in two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and proteomic profiling of mycobacterial membrane proteins remains a major challenge. Their limited solubility in aqueous buffer systems and their relatively low abundance in a background of highly abundant cytoplasmic proteins have yet to be overcome. Several studies have reported extraction of membrane- and membrane-associated proteins using centrifugation to obtain purified cell wall and cell membrane fractions for analysis by sodium-dodecyl-sulphate polyacrylamide gel electrophoresis (SDS-PAGE) in combination with liquid chromatography tandem mass spectrometry (LC-MS/MS) [10–13]. Common for these studies is pre-isolation of the membrane and cell wall of the bacteria, and application of different washing techniques prior to protein extraction by detergents.