共查询到20条相似文献,搜索用时 15 毫秒
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D Sompolinsky J B Hertz N H?iby K Jensen B Mansa V B Pedersen Z Samra 《Acta pathologica et microbiologica Scandinavica. Section B, Microbiology》1980,88(5):253-260
Common Antigen (CA) of Pseudomonas aeruginosa has been shown to be a protein composed of polypeptide subunits of a molecular weight (MW) of about 62 000. The MW of this protein was estimated to 665 000 by gel filtration on sepharose CL-6B, to 800 000 by electrophoresis on polyacrylamide gradient gels and to about 900 000 by ultracentrifugation, on a sucrose gradient. By analytical ultracentrifugation with Schlieren optics a sedimentation coefficient (S20 degrees, W) of 22.65 was calculated. The isoelectrical point was determined to pH 4.4. The antigen was decomposed on exposure to proteolytic enzymes. Polysaccharide, lipid, deoxyribonucleic acid or ribonucleic acid were not demonstrated in CA. The amino acid content of CA was determined, and no hexosamine or abnormal residues were observed. The amino acid content of CA was determined, and no hexosamine or abnormal residues were observed. The antigen was degraded when heated to 100 degrees C for 4 min or when exposed to pH below 4 or above 11 at 4 degree C. CA has been isolated from the cytoplasmic water-soluble fraction of disintegrated bacteria and only trace-amounts could be obtained from envelope fractions after solubilization with Triton X-100. 相似文献
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A serological study with monoclonal antibodies to an antigen common to a wide range of bacteria 总被引:1,自引:0,他引:1
K. Jensen J.B. Hertz J. Samra C. Magiar F.S. Larsen J. Selmer D. Sompolinsky 《FEMS microbiology letters》1985,29(1-2):129-133
Abstract Three monoclonal antibodies (MCA) against Pseudomonas aeruginosa common antigen (CA) and one MCA against Bordetella pertussis CA were produced. These immunoglobulins were examined by ELISA against extracts of a wide range of Gram-positive and Gram-negative bacteria. No reactions were obtained with Gram-positive organisms. The reaction patterns with Gram-negative organisms were different for each of the monoclonal antibodies and did not follow the accepted taxonomal principles. The results prove that a number of antigen determinants are not shared by CA of different bacteria. 相似文献
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Purification and characterization of a protein antigen from Leptospira interrogans serovar hardjo, common to a wide range of bacteria 总被引:1,自引:0,他引:1
A protein with a molecular mass of 64 kDa (P64) from Leptospira interrogans serovar hardjo was partially purified by using successively, phase partitioning with Triton X-114, ion-exchange chromatography and sucrose gradient centrifugation. Purification to homogeneity was obtained by electroelution of P64 from SDS-polyacrylamide gels. Monospecific rabbit antiserum (R alpha P64) was prepared using the purified protein preparation. P64 had a native molecular mass of greater than 670 kDa and was recognized by R alpha P64 as well as by human antisera. Western blotting of leptospiral serovars and 18 other bacterial species with R alpha P64 showed that P64 was cross-reactive with an equivalent antigen in a wide range of bacteria, indicating that it belongs to a family of antigens previously designated 'common antigen'. This putative common antigen from Leptospira appears to have a sub-surface location, but its function is not yet known. 相似文献
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Detection of common polysaccharide antigen of Pseudomonas aeruginosa with O-antiserum to Pseudomonas cerasi 总被引:1,自引:0,他引:1
Nina A. Kocharova Yuriy A. Knirel Anna Romanowska Elzbieta Romanowska Tatyana A. Makarenko Liliya S. Edvabnaya Evgeny S. Stanislavsky Galina M. Zdorovenko Lyudmila M. Yakovleva 《FEMS microbiology letters》1991,76(2):69-74
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W Scharmann 《Journal of general microbiology》1976,93(2):283-291
A toxic substance, which destroyed leucocytes from man but was inactive against erythrocytes, was demonstrated in cultures of four out of 110 strains of Pseudomonas aeruginosa tested. The toxin, designated 'leucocidin', was cell-bound as a precursor toxin, exhibiting little or no toxicity. It was converted into toxin with maximum activity by various proteases including an endogenous elastase. The production of leucocidin was directly proportional to the number of bacteria and was not influenced by variations in media, iron concentration, pH or temperature. The best method for large-scale production of leucocidin was autolysis of washed bacteria. 相似文献
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M. D. Lilly P. H. Clarke M. Houldsworth J. A. Currie P. Dunnill 《Biotechnology and bioengineering》1969,11(3):283-292
A pilot-scale process for the isolation of an aliphatic, amidase from Pseudomonas aeruginosa has been developed. A constitutive, partially irrepressible mutant was employed to give a high initial enzyme concentration. An existing laboratory isolation procedure has been scaled up and modified particularly by substitution of polyethylene glycol for ammonium sulfate precipitation as the first stage in the conversion of the fractionation to continuous operation. Full recovery of activity was achieved with the modification. The recovery of enzyme from a subsequent chromatographic stage was 85% and the maximum overall purification was 28-fold. 相似文献
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Tatyana A. Makarenko Nina A. Kocharova Lilia S. Edvabnaya Yuriy E. Tsvetkov Elena V. Kholodkova Yuriy A. Knirel Leon V. Backinowsky Nikolay K. Kochetkov Evgeny S. Stanislavsky 《FEMS immunology and medical microbiology》1993,7(3):251-256
Abstract Synthetic d -rhamnan, with the structure of Pseudomonas aeruginosa common polysaccharide antigen (CPA), was conjugated with BSA. The artificial antigen obtained, and the natural antigens, lipopolysaccharides (LPS) of P. aeruginosa and Pseudomonas cerasi with rhamnan chains of the same structure, were studied by ELISA with rabbit antibodies to the d -rhamnan-BSA conjugate and to the P. cerasi O-antigen. Immunological relations between the LPS of P. aeruginosa and P. cerasi determined by CPA as well as between these LPS and d -rhamnan-BSA were revealed by ELISA. O-antiserum to P. cerasi possesses protective activity in the mouse passive protection test when mice are challenged with some P. aeruginosa strains; the antiserum to the d -rhamnan-BSA does not possess protective activity in mice. 相似文献
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Results of minimal inhibitory concentration tests with a diversity of bacterial strains showed that 9-chloro-9-(4-diethylaminophenyl)-10-phenylacridan (C-390) inhibited the growth of all microorganisms tested (other than Pseudomonas aeruginosa) at 25 microgram/ml or less, whereas MICs obtained for P. aeruginosa ranged from to to greater than 100 microgram/ml. Therefore, C-390 was evaluated as a potential selective agent for P. aeruginosa in pseudomonas agar F. Recovery tests were conducted on this medium with 53 strains o P. aeruginosa, and the results were compared to those obtained in similar tests on commercially available selective media, i.e., pseudomonas isolation agar and Pseudosel agar. The results of these comparisons indicated that pseudomonas agar F with C-390 was significantly less inhibitory than Pseudosel agar and pseudomonas isolation agar and more selective than pseudomonas isolation agar. The incorporation of C-390 in pseudomonas agar F also provided a medium that was both selective and differential. Preliminary evidence also suggested that C-390 may be added to other basal media with comparable results. 相似文献
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OEP, a component consisting mainly of protein with small amounts of lipids and sugars, has been isolated from the autolysate of Pseudomonas aeruginosa and purified by physicochemical methods. It possesses remarkable biological properties, showing antitumor and interferon-inducing activities. As regards the antitumor activity of the sample, the ED50 value against ascites sarcoma-180 was 1 microgram/kg mouse/day, and its interferon-inducing activity amounted to 15 units at a concentration of 0.01 microgram/ml. Both activities increased after protease digestion, reaching about ten times those of the sample which had not undergone digestion. The protease-treated OEP contained 17% protein, 14.5% total sugars, 31% lipids, 12.5% hexosamine, 3.8% KDO, and 2.7% phosphorus. Neutral sugars consisted of 12.4% rhamnose, 2.7% mannose, 66.9% glucose, and other unidentified material. Total lipids derived from OEP consisted of 65% loosely-bound and 35% covalently-bound lipids; the former contained C14:10, C16:0, C16:1, C18:0, and C15:1 acids and the latter, beta-OH C10:0, C12:0, alpha-OH C12:0, beta-OH C12:0, C16:0, and C16:1 acids. The antitumor and interferon-inducing activities of OEP remained after the removal of loosely-bound lipids from OEP. 相似文献
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Melamed Jacob Kocev Alexander Torgov Vladimir Veselovsky Vladimir Brockhausen Inka 《Glycoconjugate journal》2022,39(3):393-411
Glycoconjugate Journal - The Gram-negative bacterium Pseudomonas aeruginosa simultaneously expresses two O-antigenic glycoforms. While the O-specific antigen (OSA) is variable in composition, the... 相似文献
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Thompson SE Smith M Wilkinson MC Peek K 《Applied and environmental microbiology》2001,67(9):4001-4008
A chitinase antigen has been identified in Pseudomonas aeruginosa strain 385 using sera from animals immunized with a whole-cell vaccine. The majority of the activity was shown to be in the cytoplasm, with some activity in the membrane fraction. The chitinase was not secreted into the culture medium. Purification of the enzyme was achieved by exploiting its binding to crab shell chitin. The purified enzyme had a molecular mass of 58 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and a pI of 5.2. NH2-terminal amino acid sequencing revealed two sequences of M(I/L)RID and (Q/M/V)AREDAAAAM that gave an exact match to sequences in a translated putative open reading frame from the P. aeruginosa genome. The chitinase was active against chitin azure, ethylene glycol chitin, and colloidal chitin. It did not display any lysozyme activity. Using synthetic 4-methylumbelliferyl chitin substrates, it was shown to be an endochitinase. The Km and kcat for 4-nitrophenyl-beta-D-N,N'-diacetylchitobiose were 4.28 mM and 1.7 s(-1) respectively, and for 4-nitrophenyl-beta-D-N,N',N"-triacetylchitotriose, they were 0.48 mM and 0.16 s(-1) respectively. The pH optimum was determined to be pH 6.75, and 90% activity was maintained over the pH range 6.5 to 7.1. The enzyme was stable over the pH range 5 to 10 for 3 h and to temperatures up to 50 degrees C for 30 min. The chitinase bound strongly to chitin, chitin azure, colloidal chitin, lichenan, and cellulose but poorly to chitosan, xylan, and heparin. It is suggested that the chitinase functions primarily as a chitobiosidase, removing chitobiose from the nonreducing ends of chitin and chitin oligosaccharides. 相似文献
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Monoclonal antibodies (mAbs) against the Yersinia enterocolitica immunodominant 60 kDa antigen, termed cross-reacting protein antigen (CRPA), were obtained by fusion of spleen cells from mice immunized with CRPA with murine myeloma cells. The reactivities of the mAbs were examined by Western blotting against extracts of Y. enterocolitica and 23 other species of Gram-positive and Gram-negative bacteria. Cross-reactions were recognized with a wide range of bacteria, but not with Gram-positive cocci. The reactivities were different for each mAb, suggesting that both species-specific and multiple cross-reactive epitopes were present on the CRPA molecule. CRPA was produced under heat-shock conditions in Y. enterocolitica and was shown to correspond immunologically to the GroEL protein in Escherichia coli, a protein involved in the morphogenesis of coliphage. In addition to CRPA, at least nine other major heat-shock proteins were detected by two-dimensional gel electrophoresis of extracts of heat-shocked Y. enterocolitica. 相似文献
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Leid JG Willson CJ Shirtliff ME Hassett DJ Parsek MR Jeffers AK 《Journal of immunology (Baltimore, Md. : 1950)》2005,175(11):7512-7518
The ability of Pseudomonas aeruginosa to form biofilms and cause chronic infections in the lungs of cystic fibrosis patients is well documented. Numerous studies have revealed that P. aeruginosa biofilms are highly refractory to antibiotics. However, dramatically fewer studies have addressed P. aeruginosa biofilm resistance to the host's immune system. In planktonic, unattached (nonbiofilm) P. aeruginosa, the exopolysaccharide alginate provides protection against a variety of host factors yet the role of alginate in protection of biofilm bacteria is unclear. To address this issue, we tested wild-type strains PAO1, PA14, the mucoid cystic fibrosis isolate, FRD1 (mucA22+), and the respective isogenic mutants which lacked the ability to produce alginate, for their susceptibility to human leukocytes in the presence and absence of IFN-gamma. Human leukocytes, in the presence of recombinant human IFN-gamma, killed biofilm bacteria lacking alginate after a 4-h challenge at 37 degrees C. Bacterial killing was dependent on the presence of IFN-gamma. Killing of the alginate-negative biofilm bacteria was mediated through mononuclear cell phagocytosis since treatment with cytochalasin B, which prevents actin polymerization, inhibited leukocyte-specific bacterial killing. By direct microscopic observation, phagocytosis of alginate-negative biofilm bacteria was significantly increased in the presence of IFN-gamma vs all other treatments. Addition of exogenous, purified alginate to the alginate-negative biofilms restored resistance to human leukocyte killing. Our results suggest that although alginate may not play a significant role in bacterial attachment, biofilm development, and formation, it may play an important role in protecting mucoid P. aeruginosa biofilm bacteria from the human immune system. 相似文献
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Matthew T.G. Holden Siri Ram Chhabra Rocky de Nys Paul Stead Nigel J. Bainton Philip J. Hill Mike Manefield Naresh Kumar Maurice Labatte Dacre England Scott Rice Mike Givskov George P.C. Salmond Gordon S.A.B. Stewart † Barrie W. Bycroft Staffan Kjelleberg & Paul Williams 《Molecular microbiology》1999,33(6):1254-1266
In cell-free Pseudomonas aeruginosa culture supernatants, we identified two compounds capable of activating an N-acylhomoserine lactone (AHL) biosensor. Mass spectrometry and NMR spectroscopy revealed that these compounds were not AHLs but the diketopiperazines (DKPs), cyclo(DeltaAla-L-Val) and cyclo(L-Pro-L-Tyr) respectively. These compounds were also found in cell-free supernatants from Proteus mirabilis, Citrobacter freundii and Enterobacter agglomerans [cyclo(DeltaAla-L-Val) only]. Although both DKPs were absent from Pseudomonas fluorescens and Pseudomonas alcaligenes, we isolated, from both pseudomonads, a third DKP, which was chemically characterized as cyclo(L-Phe-L-Pro). Dose-response curves using a LuxR-based AHL biosensor indicated that cyclo(DeltaAla-L-Val), cyclo(L-Pro-L-Tyr) and cyclo(L-Phe-L-Pro) activate the biosensor in a concentration-dependent manner, albeit at much higher concentrations than the natural activator N-(3-oxohexanoyl)-L-homoserine lactone (3-oxo-C6-HSL). Competition studies showed that cyclo(DeltaAla-L-Val), cyclo(L-Pro-L-Tyr) and cyclo(L-Phe-L-Pro) antagonize the 3-oxo-C6-HSL-mediated induction of bioluminescence, suggesting that these DKPs may compete for the same LuxR-binding site. Similarly, DKPs were found to be capable of activating or antagonizing other LuxR-based quorum-sensing systems, such as the N-butanoylhomoserine lactone-dependent swarming motility of Serratia liquefaciens. Although the physiological role of these DKPs has yet to be established, their activity suggests the existence of cross talk among bacterial signalling systems. 相似文献
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