doi: 10.1111/2049-632X.12033, Gerard-Vincent, M., Robert, V., Ball, G., Bleves, S., Michel, G. P., Lazdunski, A., et al. Refined crystallographic structure of Pseudomonas aeruginosa exotoxin A and its implications for the molecular mechanism of toxicity. Concentrated semi-purified exotoxin A was examined for presence of exotoxin A using the counter immunoelectrophoresis (CIEP) method. Importance of the glutamate residue of KDEL in increasing the cytotoxicity of Pseudomonas exotoxin derivatives and for increased binding to the KDEL receptor. Moreover, the molecule can be cleaved by furin, presumably to facilitate subsequent trafficking. doi: 10.1016/j.intimp.2007.05.001. Nucleic Acids Res. Protective effect of DNA vaccine encoding pseudomonas exotoxin A and PcrV against acute pulmonary P. aeruginosa Infection. tabaci 6605 (Pta6605) is a causal agent of wildfire disease in host tobacco plants. These bacilli are found everywhere and they remain in environment for several years due to their ability in producing spores. The opportunistic pathogen Pseudomonas aeruginosa (Pa) causes severe nosocomial infections, especially in immunocompromised individuals and the elderly. Three of the immunized mice (6.3%) died. Biol. Rab proteins are highly compartmentalized GTPases in organelle membranes. doi: 10.1016/j.ijmm.2008.08.003, Yates, S. P., and Merrill, A. R. (2004). 2009 Mar;299(3):161-76. doi: 10.1016/j.ijmm.2008.08.003. Manchester, England, United Kingdom. 75 white out-bred mice were provided from the Laboratory Animal Research Center of the Shiraz University of Medical Sciences, housed in an ambient temperature of 21 2C and relative humidity of 6570%, and given a balanced diet with free access to food and water. 1976, 14: 942-947. Natl. In the non-immunized mice, the colony count increased for 6 days post-inoculation with P. aeruginosa and the majority of the mice (80%) died within this period. 61, 6064. Sci. 264, 1425614261. Antitoxin and exotoxin A were detected in the sera of the experimental group by CIEP. Biotechnol Lett. This belief is founded on the demonstrable toxicity of purified preparations, and on detection of the toxin in the tissues of burned animals infected with Ps. Our results demonstrate that in a mouse model of bacterial infection in burn wounds, active immunization with semipurified exotoxin A protected against infection with P. aeruginosa and reduced mortality. To determine more precisely the ranges of immunity in the vaccinated mice, the titer of anti-exotoxin A was measured by enzyme-linked immunosorbent assay (ELISA) as previously described [14]. Reed L, Muench HA: Simple method for estimating 50% end point. Pseudomonas is a clinically significant and opportunistic pathogen, often causing nosocomial infections. Motsumoto T, Tateda K, Furuya N, Miyazaki S, Ohno A, Ishii Y, Hirakata Y, Yamaguchi K: Efficacies of alkaline protease, elastase and exotoxin A toxoid vaccines against gut-derived Pseudomonas aeruginosa sepsis in mice. See this image and copyright information in PMC. With medical big data and AI algorithms, eHealthMe . 10.1007/s00418-013-1130-9 The PE gene was originally cloned from the P. aeruginosa strain PA 103 and analysis of the 5 and 3 flanking regions evidenced that the PE gene is translated from a monocystronic message (Gray et al., 1984). The mortality rate is higher than bacteremias caused by other gram-negative opportunistic pathogens. Nature 436, 979984. 267, 1242012423. Article We demonstrated how P. aeruginosa may simultaneously develop resistance and compromise the activity of new -lactam/-lactamase inhibitor combinations when exposed to ceftazidime/avibactam through selection of mutations leading to PDC modification and up-regulation of MexAB-OprM-mediated efflux. Regulation of Golgi signaling and trafficking by the KDEL receptor. After 2-ketogluconate binding, PtxS dissociates from the PtxR/DNA complex and PtxR can recruit RNA polymerase to facilitate the transcription of the toxin (Daddaoua et al., 2012, 2014; Figure 1B). Therapeutic targeting of HCMV-encoded chemokine receptor US28: Progress and challenges. 112(Pt 4), 467475. On the TGN, the C-terminal REDL motif of PE (aa 609612) binds to the KDEL receptor and the toxin is transported to the ER in a retrograde manner (Kreitman and Pastan, 1995; Jackson et al., 1999). U.S.A. 90, 77747778. Bacillus cereus causes food poisoning and sometimes eye infections and other localized . (6 cases [14%]), Pseudomonas (5 cases [11%]), and Clostridium (4 . The mice were followed for 30 days. P. aeruginosa PA103, which produced EXA, was 20 times more virulent for normal mice than was its EXA-deficient mutant, PA103-29. The survival rate in both groups was compared. Global regulatory pathways and cross-talk control pseudomonas aeruginosa environmental lifestyle and virulence phenotype. The characterization of released extracellular vesicles/exosomes and the . Natl. After an initial colonization phase, mostly dependent on cell-associated virulence factors, the infectious process The efficacy of vaccine was calculated as the percentage survival during the 70-day observation period following inoculation with toxogenic P. aeruginosa (PA 103). 3 patients developed fever, chils, and hypotension following cardiac catheterization. Pseudomonas is a type of bacteria (germ) that is found commonly in the environment, like in soil and in water. PubMed Taken together, PE represents a remarkable molecule, which provides deep insight into pathoadaptive processes. Over this time, bacteria developed a wide spectrum of adaptation to optimize infection and survival. Pseudomonas aeruginosa adapts its iron uptake strategies in function of the type of infections. In the last years, the cytotoxic pathways of PE in eukaryotic host cells were investigated. 2004, 30: 241-243. doi: 10.1006/jmbi.2001.5195. 2002, 29: 227-. Antibodies to exotoxin A provide . To determine the effect of Pseudomonas aeruginosa exotoxin A (P-ExA) on cytokine production, we studied cytokine release induced by heat-killed P. aeruginosa (HKPA) in human whole blood in the presence or absence of P-ExA. Acad. J. Clin. Pseudomonas aeruginosa is an environmentally ubiquitous gram-negative bacterium. Biol. Chonghua LI, Nicolau DP, Lister PD, Quintiliani R, Nightingale CH: Pharmacodynamic study of B-lactamase alone and in combination with B-lactamase inhibitors against Pseudomonas aeruginosa processing an inducible b-lactamase. The intoxication pathways of PE are not fully elucidated yet. Preclinical and clinical trials with PE-based immunotoxins were reviewed elsewhere (Wolf and Elsasser-Beile, 2009; Weidle et al., 2014). No use, distribution or reproduction is permitted which does not comply with these terms. Background Pseudomonas aeruginosa is the leading cause of nosocomial infections, especially in people with a compromised immune system. Chem. 47, 5764. The ADP-ribosylation mechanism of PE was studied in detail and it turned out that it follows an SN1 nucleophilic substitution mechanism (Beattie et al., 1996; Armstrong et al., 2002; Jorgensen et al., 2005; Figure 2). Once secreted, the terminal lysine (aa 613) of PE can be cleaved from the toxin in the extracellular environment, presumably by plasma carboxypeptidases of the host. EXA was detected in the pla (1999). Exotoxin A from Pseudomonas aeruginosa is the highly toxic virulence component that bind to specific cell receptors. As expected, no exotoxin A was detected in the sera by CIEP, which may be due to neutralization of the toxin by previously antitoxins formed following immunization. Proteolytic activation of bacterial toxins: role of bacterial and host cell proteases. In the hydrophilic environment of the periplasm, PE is folded to a mature conformational protein in a manner that can be recognized by the type II secretion system (T2SS), specifically called Xcp in P. aeruginosa, for secretion into the extracellular space (Voulhoux et al., 2000; Gerard-Vincent et al., 2002). Edited by: Mandell GL, Bennettje-Dolin R. 2000, Philadelphia, PA: Churchill Livingstone, 2310-5. 8600 Rockville Pike Then a Rab6-controlled lipid-dependent sorting pathway is used for trafficking to the ER (White et al., 1999; Smith et al., 2006). J Antimicrobiol Chemother. PubMed The antimicrobial activity of silver-coated carbon nanotubes (AgCNTs) and their potential mode of action against mucoid and nonmucoid strains of Pseudomonas aeruginosa was investigated in vitro. In the absence of 2-ketogluconate, two PtxS molecules are bound to a dimer of the regulator PtxR, which again binds to the 35 region to the PE promotor and inhibits the transcription of PE. Bacterial infection is a major complication after thermal injury, especially in developing countries [1618]. CAS Cytotoxic activity of chimeric proteins composed of acidic fibroblast growth factor and Pseudomonas exotoxin on a variety of cell types. Acad. Curr. Provided by the Springer Nature SharedIt content-sharing initiative. Privacy Moreover, PE represents a remarkable example for pathoadaptive evolution, how bacterial molecules have been structurally and functionally optimized under evolutionary pressure to effectively impair and kill their host cells. Infect. Bang R, Sharma PNM, Sanyal SC, Al-najjadah I: Septicemia after burn injury: a comparative study. Anyone you share the following link with will be able to read this content: Sorry, a shareable link is not currently available for this article. 50 mice were immunized with the toxoid, burned with hot metal and infected with 1 108 . The pathogenic bacterium Pseudomonas aeruginosa has the ability to cause severe acute and chronic infections in humans. Accessibility Proc. However, Matsumato et al. This site needs JavaScript to work properly. Development of point of care medical devices for detection of white blood cells in peritonitis and antibiotic resistance in UTI's. Microbiology techniques, mammalian cell culture, clinical samples (collection, processing and analysis), in vitro assay . After cleavage in the EE, the 37 kDa PE fragment can reach the TGN by a pathway, which was shown to be independent from Rab9. Three conserved consensus sequences identify the NAD-binding site of ADP-ribosylating enzymes, expressed by eukaryotes, bacteria and T-even bacteriophages. Detection of virulence factors of Pseudomonas aeruginosa in different animals by using bacteriological and molecular methods Online ahead of print. Proteolysis of Pseudomonas exotoxin A within hepatic endosomes by cathepsins B and D produces fragments displaying in vitro ADP-ribosylating and apoptotic effects. The pathogenic bacterium Pseudomonas aeruginosa has the ability to cause severe acute and chronic infections in humans. All experiments were carried out under aseptic conditions. 48, 515521. AM plastic surgeon, main researcher, cooperated in inducing burns. Each injection contained 100 g of semi-purified toxoid in 2 mL of PBS. Knowledge about its intoxication pathways should not only be used for the construction of PE-based immunotoxins for targeted therapy (Wolf and Elsasser-Beile, 2009; Weidle et al., 2014), but also for the development of strategies to alleviate P. aeruginosa infections (Jiang et al., 2014; Farajnia et al., 2015). 2005, 31: 306-309. Sci. Microbiol. The role of exotoxin A in the mortality of experimentally-infected animals has been demonstrated [6] and the LD50 of the exotoxin reported to be 6080 ng/mouse [7]. Exotoxin. 75% of deaths following burns are related to microbial infections [19]. J Med Microbiol. doi: 10.1021/bi991308+, Nguyen, D., and Singh, P. K. (2006). Cell Biol. After cleavage and transport into late endosomes, the 37 kDa PE fragment exploits a Rab9-regulated pathway to reach the trans Golgi network (TGN). Chen TY, Lin CP, Loa CC, Chen TL, Shang HF, Hwang J, Hui CF. Genet. GtrS and GltR form a two-component system: the central role of 2-ketogluconate in the expression of exotoxin A and glucose catabolic enzymes in Pseudomonas aeruginosa. A group of 4 rabbits were immunized with the toxoid. Iglewski, B. H., Liu, P. V., and Kabat, D. (1977). 2007, 28 (6): 899-903. Export of antigenic peptides from the endoplasmic reticulum intersects with retrograde protein translocation through the Sec61p channel. Mol. On the path to uncover the bacterial type II secretion system. The ADP-ribose group is subsequently transferred to the N3 atom of the diphthamide imidazole ring, which results in the ADP-ribosylated eEF-2 protein (Armstrong et al., 2002; Jorgensen et al., 2005). Pseudomonas aeruginosa is a Gram-negative pathogen that has become an important cause of infection in humans and can be associated with significant morbidity and mortality.. Animal selection, all experiments, subsequent care and the sacrifice procedure were all performed according to the guidelines and under the supervision of the Animal Care Committee of the Iran Veterinary Organization. Active substance / international non-proprietary name (INN) / common name . Insights into diphthamide, key diphtheria toxin effector. doi: 10.1038/nature03871, Koopmann, J. O., Albring, J., Huter, E., Bulbuc, N., Spee, P., Neefjes, J., et al. Mutagenesis experiments gave evidence that two N-terminal glutamic acid residues at the +2 and +3 positions of domain Ia as well as domain II of PE are important for folding and extracellular secretion (Lu et al., 1993). 1993, 4: 345-8. The epidemiology, pathogenesis and treatment of Pseudomonas aeruginosa infections. ADP-ribosylation of eEF-2. PE is expressed as a protein with a length of 638 amino acids (aa) and can be divided into several structural and functional domains (Wedekind et al., 2001; Figure 1A). As a consequence, apoptosis is induced and the host cell irreversibly dies. doi: 10.1016/S1074-7613(00)00013-3. Biol. Targets 16, 859873. Chem. Pathog. Burns. Crit. BMC Microbiol 9, 23 (2009). Characterization of competitive inhibitors for the transferase activity of Pseudomonas aeruginosa exotoxin A. J. Pseudomonas aeruginosa produces a large number of extracellular toxins, which include phytotoxic factor, pigments, hydrocyanic acid, proteolytic enzymes, phos-pholipase, enterotoxin, exotoxin, and slime. The bacterium Pseudomonas aeruginosa is a dangerous and opportunistic bacteria responsible for severe nosocomial infections, and chronic infections in cystic fibrosis patients. Initially, the PE-fragment binds to NAD+ and interacts via the so-called active-site loop L4 (aa 483490 of domain III) with eEF-2 (Yates and Merrill, 2004). 1996, 44: 145-153. A toxigenic strain of P. aeruginosa (PA 103) was used for exotoxin A preparation. The LD50 was determined according to the Reed and Muench method [13] and calculated to be 0.5 g. No report of Pseudomonas infection is found in people who take Sarisol no. N. Engl. The remainder (48 mice) were challenged with a lethal dose of P. aeruginosa and followed for 70 days. 2-ketogluconate is able to bind to the transcriptional repressor protein PtxS. With regard to its function it is specified as NAD+-diphthamide-ADP-ribosyltransferase (EC 2.4.2.36) (Domenighini and Rappuoli, 1996). 147, 743760. Immun. doi: 10.1042/BJ20031731, Zerial, M., and McBride, H. (2001). 10.1021/bi961985t (2013). Exotoxin A is currently thought to be the principal lethal factor in experimental Pseudomonas aeruginosa infection. Translocation through the Sec61p channel developing countries [ 1618 ] ) died for severe infections! Researcher, cooperated in inducing burns for normal mice than was its mutant. 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