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Bacterial Cell Wall (bacterial + cell_wall)
Selected AbstractsNod1 and Nod2 induce CCL5/RANTES through the NF-,B pathwayEUROPEAN JOURNAL OF IMMUNOLOGY, Issue 9 2007Catherine Werts Abstract The Nod-like receptor proteins Nod1 and Nod2 participate in innate immune responses against bacteria through intracellular detection of peptidoglycan, a component of bacterial cell wall. Recent evidence has demonstrated that Nod1 stimulates the release of chemokines that attract neutrophils at the site of infection, such as CXCL8/IL-8 in humans, and CXCL1/keratinocyte-derived chemokine and CXCL2/MIP-2 in mice. We aimed to determine whether Nod proteins could trigger the release of CCL5/RANTES, a chemokine known to attract a number of immune cells, but not neutrophils. Our results demonstrate that activation of both Nod1 and Nod2 results in substantial secretion of CCL5 by murine macrophages. Moreover, in vivo, the intraperitoneal injection of murine Nod1 or Nod2 agonists resulted in a rapid secretion of CCL5 into the bloodstream. We also observed that Nod-dependent secretion of CCL5 did not correlate with the induction of the interferon-, pathway, a major signaling cascade for the activation of CCL5 by viruses. In contrast, we identified a key role of the NF-,B pathway in Nod-dependent stimulation of the CCL5 promoter. Together, these results identify a novel target downstream of Nod1 and Nod2, which is likely to play a key role in orchestrating the global Nod-dependent immune defense during bacterial infections. [source] Cell wall growth during elongation and division: one ring to bind them?MOLECULAR MICROBIOLOGY, Issue 4 2007Dirk-Jan Scheffers Summary The role of the cell division protein FtsZ in bacterial cell wall (CW) synthesis is believed to be restricted to localizing proteins involved in the synthesis of the septal wall. In this issue of Molecular Microbiology, the groups of Christine Jacobs-Wagner and Waldemar Vollmer provide compelling evidence that in Caulobacter crescentus, FtsZ plays an additional role in CW synthesis in non-dividing cells. During elongation (cell growth) FtsZ is responsible for the incorporation of CW material in a zone at the midcell by recruiting MurG, a protein involved in peptidoglycan (PG) precursor synthesis. This resembles earlier findings of FtsZ mediated PG synthesis activity in Escherichia coli. A role of FtsZ in PG synthesis during elongation forces a rethink of the current model of CW synthesis in rod-shaped bacteria. [source] Structural analysis of an "open" form of PBP1B from Streptococcus pneumoniaePROTEIN SCIENCE, Issue 7 2006Andrew L. Lovering Abstract The class A PBP1b from Streptococcus pneumoniae is responsible for glycosyltransferase and transpeptidase (TP) reactions, forming the peptidoglycan of the bacterial cell wall. The enzyme has been produced in a stable, soluble form and undergoes time-dependent proteolysis to leave an intact TP domain. Crystals of this TP domain were obtained, diffracting to 2.2 Å resolution, and the structure was solved by using molecular replacement. Analysis of the structure revealed an "open" active site, with important conformational differences to the previously determined "closed" apoenzyme. The active-site nucleophile, Ser460, is in an orientation that allows for acylation by ,-lactams. Consistent with the productive conformation of the conserved active-site catalytic residues, adjacent loops show only minor deviation from those of known acyl-enzyme structures. These findings are discussed in the context of enzyme functionality and the possible conformational sampling of PBP1b between active and inactive states. [source] Structures of an alanine racemase from Bacillus anthracis (BA0252) in the presence and absence of (R)-1-aminoethylphosphonic acid (l -Ala-P)ACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 5 2008Kinfai Au Bacillus anthracis, the causative agent of anthrax, has been targeted by the Oxford Protein Production Facility to validate high-throughput protocols within the Structural Proteomics in Europe project. As part of this work, the structures of an alanine racemase (BA0252) in the presence and absence of the inhibitor (R)-1-aminoethylphosphonic acid (l -Ala-P) have determined by X-ray crystallography to resolutions of 2.1 and 1.47,Å, respectively. Difficulties in crystallizing this protein were overcome by the use of reductive methylation. Alanine racemase has attracted much interest as a possible target for anti-anthrax drugs: not only is d -alanine a vital component of the bacterial cell wall, but recent studies also indicate that alanine racemase, which is accessible in the exosporium, plays a key role in inhibition of germination in B. anthracis. These structures confirm the binding mode of l -Ala-P but suggest an unexpected mechanism of inhibition of alanine racemase by this compound and could provide a basis for the design of improved alanine racemase inhibitors with potential as anti-anthrax therapies. [source] Specific Labeling of Peptidoglycan Precursors as a Tool for Bacterial Cell Wall StudiesCHEMBIOCHEM, Issue 4 2009Vincent van Dam Abstract Wall chart: The predominant component of the bacterial cell wall, peptidoglycan, consists of long alternating stretches of aminosugar subunits interlinked in a large three-dimensional network and is formed from precursors through several cytosolic and membrane-bound steps. The high tolerance of the cell wall synthesis machinery allows for the use of labeled precursor derivatives to study diverse aspects of bacterial cell wall synthesis and interaction with antibiotics. Because of its importance for bacterial cell survival, the bacterial cell wall is an attractive target for new antibiotics in a time of great demand for new antibiotic compounds. Therefore, more knowledge about the diverse processes related to bacterial cell wall synthesis is needed. The cell wall is located on the exterior of the cell and consists mainly of peptidoglycan, a large macromolecule built up from a three-dimensional network of aminosugar strands interlinked with peptide bridges. The subunits of peptidoglycan are synthesized inside the cell before they are transported to the exterior in order to be incorporated into the growing peptidoglycan. The high flexibility of the cell wall synthesis machinery towards unnatural derivatives of these subunits enables research on the bacterial cell wall using labeled compounds. This review highlights the high potential of labeled cell wall precursors in various areas of cell wall research. Labeled precursors can be used in investigating direct cell wall,antibiotic interactions and in cell wall synthesis and localization studies. Moreover, these compounds can provide a powerful tool in the elucidation of the cell wall proteome, the "wallosome," and thus, might provide new targets for antibiotics. [source] Structure-based Optimization of MurF InhibitorsCHEMICAL BIOLOGY & DRUG DESIGN, Issue 1 2006Geoffrey F. Stamper The d -Ala- d -Ala adding enzyme (MurF) from Streptococcus pneumoniae catalyzes the ATP-dependent formation of the UDP-MurNAc-pentapeptide, a critical component of the bacterial cell wall. MurF is a potential target for antibacterial design because it is unique to bacteria and performs an essential non-redundant function in the bacterial cell. The recent discovery and subsequent cocrystal structure determination of MurF in complex with a new class of inhibitors served as a catalyst to begin a medicinal chemistry program aimed at improving their potency. We report here a multidisciplinary approach to this effort that allowed for rapid generation of cocrystal structures, thereby providing the crystallographic information critical for driving the inhibitor optimization process. This effort resulted in the discovery of low-nanomolar inhibitors of this bacterial enzyme. [source] Conserved cytoplasmic motifs that distinguish sub-groups of the polyprenol phosphate:N -acetylhexosamine-1-phosphate transferase familyFEMS MICROBIOLOGY LETTERS, Issue 2 2000Matt S. Anderson Abstract WecA, MraY and WbcO are conserved members of the polyprenol phosphate:N -acetylhexosamine-1-phosphate transferase family involved in the assembly of bacterial cell walls, and catalyze reactions involving a membrane-associated polyprenol phosphate acceptor substrate and a cytoplasmically located UDP- D -amino sugar donor. MraY, WbcO and WecA purportedly utilize different UDP-sugars, although the molecular basis of this specificity is largely unknown. However, domain variations involved in specificity are predicted to occur on the cytoplasmic side of the membrane, adjacent to conserved domains involved in the mechanistic activity, and with access to the cytoplasmically located sugar nucleotides. Conserved C-terminal domains have been identified that satisfy these criteria. Topological analyses indicate that they form the highly basic, fifth cytoplasmic loop between transmembrane regions IX and X. Four diverse loops are apparent, for MraY, WecA, WbcO and RgpG, that uniquely characterize these sub-groups of the transferase family, and a correlation is evident with the known or implied UDP-sugar specificity. [source] Phosphate buffer,extractable organic nitrogen as an index of soil-N availability for sorghum and pearl milletJOURNAL OF PLANT NUTRITION AND SOIL SCIENCE, Issue 2 2010Asako Mori Abstract The availability of soil nitrogen (N) is usually quantified by the amount of mineralized N as determined after several weeks of soil incubation. Various alternative methods using chemical solvents have been developed to extract the available organic N, which is easily mineralized. We compared one such solution, neutral phosphate buffer (NPB), with conventional incubation and 0.01 M,CaCl2 extraction, as measures of soil N available to two major cereal crops of the semiarid tropics, based on the total N uptake by plants in a pot experiment. Mineralized N had the highest correlation with N uptake by pearl millet (Pennisetum glaucum L., r = 0.979***) and sorghum (Sorghum bicolor [L.] Moench, r = 0.978***). NPB-extractable N was also highly correlated with N uptake (pearl millet, r = 0.876***; sorghum, r = 0.872***). Only one major peak was detected when NPB extracts were analyzed using size-exclusion high-performance liquid chromatography, regardless of soil properties. In addition, the organic N extracted with NPB was characterized by determining the content of peptidoglycan, the main component of bacterial cell walls. Although the characteristics of NPB-extractable organic N are still unclear, it offers a promising quick assay of available N. [source] The high-resolution structure of dihydrodipicolinate synthase from Escherichia coli bound to its first substrate, pyruvateACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 12 2008Sean R. A. Devenish Dihydrodipicolinate synthase (DHDPS) mediates the key first reaction common to the biosynthesis of (S)-lysine and meso -diaminopimelate, molecules which play a crucial cross-linking role in bacterial cell walls. An effective inhibitor of DHDPS would represent a useful antibacterial agent; despite extensive effort, a suitable inhibitor has yet to be found. In an attempt to examine the specificity of the active site of DHDPS, the enzyme was cocrystallized with the substrate analogue oxaloacetate. The resulting crystals diffracted to 2.0,Å resolution, but solution of the protein structure revealed that pyruvate was bound in the active site rather than oxaloacetic acid. Kinetic analysis confirmed that the decarboxylation of oxaloacetate was not catalysed by DHDPS and was instead a slow spontaneous chemical process. [source] | |||||||||||||