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Patterns of LPS synthesis in gram negative bacteria.

LPS Biosynthesis | Junop Lab

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LPS induces SAA synthesis from human hepatocytes.

So, how is LPS biosynthesis inhibited? A few years ago, a key enzyme – a zinc-dependent metalloamidase – in LPS biosynthesis was identified; the enzyme, produced by an essential, single copy gene, is referred to as ‘LpxC’, a.k.a. UDP-3-O-(R-3-hydroxymyristoyl)-N-acetylglucosamine deacetylase – charming name, isn’t it? – and it catalyzes the committed step of synthesis of lipid A (the lipid component of LPS) in virtually all Gram-negative bacteria []. Since then, given the role of LPS in the pathogenicity of many Gram-negative organisms, LpxC has been designated a good target for developing therapeutics for these group of microbes, and several small molecule inhibitors of this enzyme have been designed and studied. The study by Lin et al. used an investigational LpxC inhibitor that is in advanced preclinical development, called LpxC-1.

and two other genes that might be involved in the LPS synthesis.

Whereas synthesis of the lipid A component of bacterial lipopolysaccharide (LPS) occurs in a well characterized, stepwise manner, assembly of the O-polysaccharide part is less defined. Genetic studies have identified a -dependent biosynthetic pathway but practical difficulties have impeded its verification and prevented its exploitation. By chemically reconstituting the pathway, Peng George Wang and colleagues now confirm that Wzy acts as a polymerase and Wzz as a chain-length regulator. Their study, reported in , increases scope for manipulation of polysaccharide biosynthesis in the lab.

Lps synthesis | pacomsividenribidingtelldali

Patterns of LPS synthesis in Gram negative bacteria

AB - We have cloned and sequenced the rfaH gene from Salmonella enterica serovar Typhi strain Ty2. The gene showed a high degree of similarity to the rfaH genes from Escherichia coli K-12 and S. enterica serovar Typhimurium. A rfaH mutant was constructed by site-directed mutagenesis. This mutant produced a rough lipopolysaccharide (LPS), with an incomplete core region. The defect in LPS expression that results from the rfaH mutation was corrected by a plasmid carrying the intact gene. The plasmid-borne rfaH gene also restored normal LPS synthesis in a rfaH mutant of E. coli. Reverse transcription-polymerase chain reaction analyses were performed to determine the effects of various environmental conditions on the expression of rfaH. The transcription of rfaH showed a growth-phase-dependent regulation, with maximal expression at the late exponential phase. Other environmental conditions, such as temperature or medium osmolarity, did not affect transcription of rfaH.

Pseudomonas aeruginosa (PA) is an important airway pathogen of cystic fibrosis and chronic obstructive disease patients. Multiply drug resistant PA is becoming increasing prevalent and new strategies are needed to combat such insidious organisms. We have previously shown that a mucoid, mucA22 mutant PA is exquisitely sensitive to acidified nitrite (A-NO2-, pH 6.5) at concentrations that are well tolerated in humans. Here, we used a transposon mutagenesis approach to identify PA mutants that are hypersensitive to A-NO2-. Among greater than 10,000 mutants screened, we focused on PA4455, in which the transposon was found to disrupt the production of a putative cytoplasmic membrane-spanning ABC transporter permease. The PA4455 mutant was not only highly sensitive to A-NO2-, but also the membrane perturbing agent, EDTA and the antibiotics doxycycline, tigecycline, colistin, and chloramphenicol, respectively. Treatment of bacteria with A-NO2- plus EDTA, however, had the most dramatic and synergistic effect, with virtually all bacteria killed by 10 mM A-NO2-, and EDTA (1 mM, aerobic, anaerobic). Most importantly, the PA4455 mutant was also sensitive to A-NO2- in biofilms. A-NO2- sensitivity and an anaerobic growth defect was also noted in two mutants (rmlC and wbpM) that are defective in B-band LPS synthesis, potentially indicating a membrane defect in the PA4455 mutant. Finally, this study describes a gene, PA4455, that when mutated, allows for dramatic sensitivity to the potential therapeutic agent, A-NO2- as well as EDTA. Furthermore, the synergy between the two compounds could offer future benefits against antibiotic resistant PA strains.

Cytokine- and LPS-induced synthesis of interleukin-8 …

The plasmid-borne rfaH gene also restored normal LPS synthesis in a rfaH mutant of E

Activation of microglial cells was observed with obvious increasing iba1-immunoreactivity following LPS stimulation in cell culture. Synthesis and up-regulation of proNGF protein significantly occurred in N9 and BV2 cells 12h-48h after LPS exposure, whereas no significant changes of proBDNF and MMP9 were observed in these microglial cell lines with LPS insult. More interestingly, extracellular release or secretion of proNGF molecule was also detected in culture medium of N9 cells after LPS stimulation. Finally, bioassay using MTT, Hoechst/PI and TUNEL staining in SH-SY5Y cells further confirmed that proNGF treatment could result in apoptotic cell death but it did not significantly influence cell viability of SH-SY5Y cells.

This in vitro study revealed LPS-stimulated proNGF synthesis and release in activated N9/BV2 microglial cell lines, also suggesting that proNGF may appeal a new pathway or possible mechanism underlying microglial toxicity in the neuroinflammation and a potential target for therapeutic manipulation of the neurodegenerative diseases.

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  • LPS-Induced proNGF Synthesis and ..

    Our contribution to these efforts is focused on disruption of lipopolysaccharide ( LPS) biosynthesis

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    30/12/2005 · Brucella spp noncanonical LPS: structure, biosynthesis, and interaction with host immune ..

  • pylori LPS biosynthesis pathway

    27/12/2017 · On Jul 1, 1985 Fern Hunt published: Patterns of LPS synthesis in Gram negative bacteria

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This paper demonstrates that LPS synthesis in Sh54 is impaired

T1 - Synthesis of unsymmetrical monocarbonyl curcumin analogues with potent inhibition on prostaglandin E2 production in LPS-induced murine and human macrophages cell lines

Chemical Synthesis of Lipopolysaccharide Core | …

In the CNS, neurotrophins such as NGF and BDNF are initially synthesized as pro-neurotrophins, that are then cleaved to release mature C-terminal forms. The proforms of neurotrophins such as proNGF and proBDNF, preferentially bind to p75NTR-sortilin receptor whereas mature neurotrophins are preferred ligands for Trk receptors. While signals emanating from Trks support neuronal survival, cell growth and synaptic strengthening, the proNGF-p75NTR-sortilin signaling can induce apoptosis, attenuate growth and weaken synaptic signaling [–]. Accumulating evidences have indicated that p75NTR-sortilin signaling triggered by abnormality or imbalance of proNGF/NGF might be involved in the glial-neuronal interaction, degenerative loss of motor neurons or cholinergic neurons, disease onset or progression in AD, PD and ALS [–]. However, it still remains a critical question if proforms of neurotrophins can be synthesized and directly secreted from the activated glial cells. By applying N9 and BV2 cell culture and lipopolysaccharide (LPS) exposure model in this study, therefore, we examined dynamic patterns of activated microglial cells and revealed LPS-induced proNGF synthesis and release from these activated microglial cells.

Chemical Synthesis of Lipopolysaccharide Core

I hope you’ve understood the significance of that observation, gentle reader. It seems to indicate that if there is a means to stop the LPS from being shed or produced, the virulence of the A. baumannii strain may perhaps be attenuated. And that’s exactly what the authors found out: “Inhibition of LPS biosynthesis did not kill A. baumannii but enhanced opsonophagocytosis and decreased inflammation, resulting in protection of mice from lethal infection. Which means, that shutting down the production of LPS did not kill the bacteria in the traditional way (the way, say, an antibiotic works), but reduced the excessive inflammation in the host, giving a chance to host immune cells, such as neutrophils and macrophages, to destroy the bacteria.

Lipopolysaccharide signaling induces serum amyloid A …

After LPS exposure, these different cultures were subjected to immunocytochemical and western blotting detection of proNGF, NGF, proBDNF, BDNF and MMP-9 synthesis in the cells. Besides, to detect direct release of proNGF extracellularly, the 15 ml conditioned medium was also collected from LPS 24h and 48h groups, centrifuged at 800g for 5 min to precipitate non-adherent cells. The supernatant was collected and filtered through a 0.2μm membrane to remove any cell debris. The filtrate was concentrated to 500μl using Amicon Ultra-15 Centrifugal Filter Devices (Millipore). Then the 500 μl filtrate was further concentrated to about 50μl using Amicon Ultra-0.5 Centrifugal Filter Devices (Millipore). About 25μl of the final filtrate was separated by 12% SDS-PAGE and the remaining proteins were detected by western blot protocol as following.

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