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1.
Microbios ; 66(267): 83-93, 1991.
Artigo em Inglês | MEDLINE | ID: mdl-2062249

RESUMO

Lipopolysaccharides (LPS) from an opaque and a translucent colony variant of Vibrio vulnificus were isolated by several methods and the electrophoretic profiles were analysed. The phenol-water extraction method provided a better yield compared to the phenol-chloroform-petroleum ether method. In addition, two rapid micro-assays were used to isolate LPS for electrophoretic analysis. The electrophoretic pattern was the same for all LPS extracts and was similar in both variants. No high molecular weight bands, characteristic of smooth LPS, were detected in the LPS of this organism. This was in contrast to the smooth nature of the LPS of another member of the Vibrionaceae examined, V. cholerae. The result of this study showed no correlation between LPS and colony morphology in V. vulnificus.


Assuntos
Lipopolissacarídeos/análise , Vibrio/análise , Eletroforese em Gel de Poliacrilamida , Lipopolissacarídeos/isolamento & purificação , Peso Molecular
3.
Appl Environ Microbiol ; 56(6): 1926-31, 1990 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-2166473

RESUMO

A rapid particle agglutination assay (PAA) utilizing latex beads coated with connective tissue and serum proteins was evaluated for its ability to identify fibronectin, collagen (types I and IV), fibrinogen, and transferrin cell surface receptors on Vibrio and Aeromonas strains isolated from diseased fish, human infections, and the environment. Similar tests were performed to screen for cell surface lectins. Vibrio as well as Aeromonas strains were found to bind connective tissue proteins (collagen types I, II, and IV and fibronectin), serum proteins (i.e., fibrinogen), and glycoproteins (bovine submaxillary mucin, hog gastric mucin, orosomucoid, and fetuin) immobilized on the latex particles. The specificity of the agglutination reaction was studied by particle agglutination inhibition assays performed by preincubating bacterial suspensions in solutions containing either gelatin (for the various connective tissue protein PAA reagents) or sialic acid-rich glycoproteins (for the various glycoprotein PAA reagents). Expression of cell surface receptors for connective tissue proteins was found to depend on culture methods.


Assuntos
Aeromonas/análise , Receptores de Superfície Celular/análise , Receptores Imunológicos/análise , Vibrio/análise , Animais , Membrana Celular/análise , Meios de Cultura , Peixes , Humanos , Testes de Fixação do Látex , Glicoproteínas da Membrana de Plaquetas/análise , Receptores de Colágeno , Receptores de Fibronectina , Receptores da Transferrina/análise , Sensibilidade e Especificidade
4.
J Bacteriol ; 172(6): 3515-8, 1990 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-2345157

RESUMO

A high level of a trans-unsaturated fatty acid was found in the phospholipids of a psychrophilic bacterium, Vibrio sp. strain ABE-1. This fatty acid was identified as 9-trans-hexadecenoic acid (C16:19t) by gas-liquid chromatography and infrared absorption spectrometry. C16:1(9)t accounted for less than 1% of the total fatty acids in cells grown at 5 degrees C and reached 12% of the total at 20 degrees C. We suggest that the increase in the level of the trans-unsaturated fatty acid is related to the high growth rate of this bacterium at elevated temperatures. Possible biological roles of the trans-unsaturated fatty acid in the adaptation of the microorganism to the ambient temperature are discussed.


Assuntos
Ácidos Graxos Insaturados/análise , Vibrio/análise , Cromatografia Gasosa , Temperatura , Vibrio/crescimento & desenvolvimento
5.
Biochem Cell Biol ; 68(2): 547-51, 1990 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-1693085

RESUMO

Vibrio vulnificus is a marine bacterium associated with both primary septicemias and wound infections in humans. The lipopolysaccharides of a virulent and an avirulent strain of Vibrio vulnificus were compared with respect to their chemical constituents and electrophoretic characteristics. 2-Keto-3-deoxyoctonic acid, a normal constituent of the lipopolysaccharide of typical Enterobacteriaceae, was not found in the lipopolysaccharide of either strain. Hexadecenoate (C16:1) was the predominant fatty acid of the lipid A moiety of the lipopolysaccharides and of the membrane phospholipids of both strains. Hydroxy fatty acids composed 44% of the total fatty acids of the lipid A of the avirulent and 40% of those in the virulent strain. In addition, odd-numbered fatty acids were detected in both lipopolysaccharides. The electrophoretic profile was similar for both strains, but demonstrated no "ladder-like" pattern characteristic of "smooth" lipopolysaccharides. The result of this study showed no significant differences between the lipopolysaccharides of the virulent and avirulent strains of Vibrio vulnificus. The possible role for lipopolysaccharide in pathogenesis of Vibrio vulnificus infections is discussed.


Assuntos
Lipopolissacarídeos/análise , Vibrio/análise , Antígenos de Bactérias/análise , Eletroforese em Gel de Poliacrilamida , Ácidos Graxos/análise , Humanos , Antígenos O , Fosfolipídeos/análise , Sepse/etiologia , Açúcares Ácidos/análise , Vibrio/imunologia , Vibrio/patogenicidade , Vibrioses/etiologia , Virulência , Infecção dos Ferimentos/etiologia
6.
Nihon Saikingaku Zasshi ; 44(6): 805-12, 1989 Nov.
Artigo em Japonês | MEDLINE | ID: mdl-2621792

RESUMO

Vibrio anguillarum, one of the causative agents of fish vibriosis, is serologically and biochemically divided into three groups (A, B and C). The chemical composition and molecular architecture of lipopolysaccharide (LPS) isolated from V. anguillarum PT 514, which belongs to serogroup B, were investigated. The LPS contained glucose (Glc), fructose (Fru), L-glycero-D-mannoheptose (L-D Hep), glucosamine (GlcN) and 4-amino-4,6-dideoxyglucose as sugar constituents in molar ratios of 8.9:0.7:3.0:1.1:1.6. Sephadex G-50 gel-chromatography of a degraded polysaccharide fraction separated from the LPS by 5% acetic acid hydrolysis suggested that the O-specific polysaccharide region consists of, in average, as much as 29 moles of Glc per 3 moles of L-D Hep, while the core polysaccharide contains at least Glc, L-D Hep and GlcN in molar ratios of 3.2 : 3.0 : 0.2. Fru and 4-amino-4,6-dideoxyglucose components were released from LPS on weak-acid hydrolysis, indicating that PT 514 LPS is distinguishable from those of Vibrio anguillarum belonging to the other serogroups. 2-Keto-3-deoxyoctonate (KDO), a common sugar constituent of gram-negative bacterial LPS, was not detected by Weissbach's color reaction under the conventional hydrolysis condition, but O-phosphoryl KDO was found in the strong-acid hydrolysate (4 M HCl, 100 C, 45 min). This substance was identical, at least in high-voltage paper electrophoresis, to 5-O-phosphoryl KDO.(ABSTRACT TRUNCATED AT 250 WORDS)


Assuntos
Lipopolissacarídeos/análise , Vibrio/análise , Carboidratos/análise , Cromatografia em Gel , Endotoxinas , Ácidos Graxos/análise , Lipídeo A/análise , Lipopolissacarídeos/isolamento & purificação
7.
FEMS Microbiol Lett ; 52(1-2): 171-5, 1989 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-2513246

RESUMO

The characteristics of Vh-rTDH, a hemolysin similar to Vp-TDH of Vibrio parahaemolyticus produced by clinical and environmental isolates of Vibrio hollisae, were comparatively studied. All 7 strains of V. hollisae tested were found to produce indistinguishable Vh-rTDH when they were examined by heat-stability test, Western blotting analysis and conventional polyacrylamide slab gel electrophoresis.


Assuntos
Proteínas Hemolisinas/análise , Vibrio/análise , Animais , Galinhas , Microbiologia Ambiental , Proteínas Hemolisinas/biossíntese , Temperatura Alta , Humanos , Imunodifusão , Peso Molecular , Coelhos , Ovinos
8.
Int J Pept Protein Res ; 34(4): 277-86, 1989 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-2599766

RESUMO

Several proteins and polypeptides of reptilian, amphibian, insect, and microbial origin share a common cytolytic property. However, these cytolysins fulfill different objectives. They provide offensive armament in the case of toxins, but defensive systems in the case of antibacterial peptides. The sequences of several nonenzymatic cytolysins and their analogues were compared to identify the structural requirements for cytolytic activity. These cytolysins, although isolated from phylogenetically unrelated organisms, possess the common sequence features of a cationic site flanked by a hydrophobic surface. The presence of such a region apparently confers the cytolytic activity of various cytolysins. The concept of a cytolytic region is strongly supported by the existence of several natural and synthetic analogues of cytolysins and by chemical modification studies of these cytolysins. This prediction provides a new focus for cytolysin research. The understanding of this structure-function relationship should facilitate the design, synthesis, and development of better antibacterial and anticancer peptides.


Assuntos
Antibacterianos/análise , Peptídeos Catiônicos Antimicrobianos , Cardiopatias/induzido quimicamente , Proteínas Hemolisinas/análise , Proteínas de Insetos , Doenças Musculares/induzido quimicamente , Toxinas Biológicas/análise , Proteínas de Xenopus , Sequência de Aminoácidos , Toxinas Bacterianas/análise , Sobrevivência Celular/efeitos dos fármacos , Endotoxinas/análise , Cardiopatias/fisiopatologia , Hormônios de Inseto/análise , Magaininas , Meliteno/análise , Dados de Sequência Molecular , Doenças Musculares/fisiopatologia , Peptídeos/análise , Fosfolipases A/análise , Proteínas Citotóxicas Formadoras de Poros , Conformação Proteica , Venenos de Serpentes/análise , Toxinas Biológicas/toxicidade , Vibrio/análise
9.
Toxicon ; 27(4): 439-64, 1989.
Artigo em Inglês | MEDLINE | ID: mdl-2658204

RESUMO

Enzyme-linked immunosorbent assays using polyclonal and monoclonal antibodies specific for V. vulnificus cytolysin detected the toxin in an extract of skin lesions and in serum from mice showing local and systemic V. vulnificus disease after subcutaneous injection of the bacterium. The cytolysin also was detected in skin lesions by an indirect immunofluorescence procedure using polyclonal and monoclonal antibodies. Our findings provide direct evidence that the cytolysin is produced in vivo during the development of the disease process, and this observation is consistent with the hypothesis that the toxin is involved in the pathogenesis of V. vulnificus disease.


Assuntos
Citotoxinas/análise , Vibrioses/metabolismo , Vibrio/análise , Animais , Citotoxinas/imunologia , Ensaio de Imunoadsorção Enzimática , Feminino , Imunofluorescência , Técnicas In Vitro , Camundongos , Camundongos Endogâmicos BALB C , Coelhos , Pele/anatomia & histologia
10.
J Gen Microbiol ; 134(6): 1699-705, 1988 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-3221203

RESUMO

A comparative study was made of the quantitative sugar composition of lipopolysaccharides (LPS) isolated from Vibrio cholerae (O1 and non O1 groups), 'V. albensis', 'V. proteus' and V. metschnikovii. The amino sugars 4-amino-4,6-dideoxy-D-mannose (perosamine) and 2-amino-2,6-dideoxy-D-glucose (quinovosamine) were present exclusively in LPS isolated from S-form O1 group of V. cholerae regardless of serotype (i.e. Ogawa or Inaba) and biotype (i.e. classical or eltor). Classical O1 group V. cholerae was distinguishable from eltor O1 group V. cholerae on the basis of the fructose content of the LPS: greater than 3% and less than or equal to 1%, respectively. Distinct differences in the sugar composition of LPS were observed between V. cholerae and 'V. albensis', 'V. proteus' and V. metschnikovii.


Assuntos
Carboidratos/análise , Lipopolissacarídeos/metabolismo , Vibrio/metabolismo , Glucosamina/análise , Hidrólise , Fosforilação , Açúcares Ácidos/análise , Vibrio/análise , Vibrio cholerae/análise , Vibrio cholerae/metabolismo
11.
Infect Immun ; 56(4): 954-60, 1988 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-3126150

RESUMO

An extracellular cytolysin produced by Vibrio metschnikovii was purified by acid precipitation, phenyl-Sepharose CL-4B chromatography, and rechromatography on a phenyl-Sepharose CL-4B column and high-performance liquid chromatography on a Mono Q (anion-exchange) column. The purified cytolysin had a molecular weight of 50,000 and an isoelectric point of 5.1. It was inactivated by heating at 60 degrees C for 5 min and was inhibited by Zn2+, Cu2+, and high concentrations of cholesterol. Lysis of calf erythrocytes by cytolysin was temperature dependent and occurred only above 18 degrees C. Moreover, no lysis was observed at high concentrations of erythrocytes, suggesting that the cytolysin lyses erythrocytes by a multihit mechanism. This cytolysin had no immunological cross-reactivities with hemolysins from other Vibrio species tested, indicating that it is a new cytolysin. V. metschnikovii cytolysin lysed erythrocytes from several animal species (calf, rabbit, guinea pig, mouse, human, sheep, chicken, and horse) and cultured cells (Vero and Chinese hamster ovary), caused fluid accumulation in the intestines of infant mice, and increased vascular permeability in rabbit skin.


Assuntos
Citotoxinas/isolamento & purificação , Vibrio/análise , Animais , Bioensaio , Cátions Bivalentes/farmacologia , Colesterol/farmacologia , Cromatografia , Cromatografia Líquida de Alta Pressão , Ensaio de Imunoadsorção Enzimática , Hemólise , Temperatura Alta , Imunodifusão , Técnicas In Vitro , Ponto Isoelétrico , Peso Molecular , Testes de Neutralização , Azul Tripano/farmacologia
12.
Biochem Biophys Res Commun ; 146(1): 101-6, 1987 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-3606610

RESUMO

A low molecular weight protein (approximately 25,000 D) exhibiting a yellow fluorescence emission peaking at approximately 540 nm was isolated from Vibrio fischeri (strain Y-1) and purified to apparent homogeneity. FMN is the chromophore, but it exhibits marked red shifts in both the absorption (lambda max = 380, 460 nm) and the fluorescence emission. When added to purified luciferase from the same strain, which itself catalyzes an emission of blue-green light (lambda max approximately 495 nm), this protein induces a bright yellow luminescence (lambda max approximately 540 nm); this corresponds to the emission of the Y-1 strain in vivo. This yellow bioluminescence emission is thus ascribed to the interaction of these two proteins, and to the excitation of the singlet FMN bound to this fluorescent protein.


Assuntos
Vibrio/análise , Proteínas Virais/isolamento & purificação , Mononucleotídeo de Flavina , Fluorescência , Peso Molecular , Espectrometria de Fluorescência
14.
Infect Immun ; 55(3): 626-30, 1987 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-3818087

RESUMO

Vibrio hollisae, a halophilic bacterium isolated from patients with diarrhea, was examined for virulence factor production. Intragastric administration of 2 X 10(7) CFU per mouse elicited fluid accumulation which peaked at ca. 6 h postchallenge in infant mice. An enterotoxin which elongated Chinese hamster ovary (CHO) cells was detected in extracts of infected-mouse intestines and in culture fluids from various growth media. The yield of the enterotoxin was maximal beginning at the onset of the stationary phase of growth in heart infusion broth supplemented with 0.5% NaCl. A concentrated preparation obtained by ammonium sulfate precipitation of culture supernatant fluids induced intestinal fluid accumulation which peaked at 2 h postchallenge in infant mice. The abilities of the enterotoxin preparation to elongate CHO cells and to elicit fluid accumulation in infant mice were inseparable by gel filtration, isoelectric focusing, and hydrophobic interaction chromatography. The enterotoxin has a molecular weight of ca. 33,000 by gel filtration and an isoelectric point of ca. 4 and is sensitive to heat.


Assuntos
Toxinas Bacterianas/isolamento & purificação , Enterotoxinas/isolamento & purificação , Conteúdo Gastrointestinal/análise , Vibrio/análise , Animais , Toxinas Bacterianas/farmacologia , Líquidos Corporais/análise , Linhagem Celular , Cromatografia em Gel , Cricetinae , Cricetulus , Enterotoxinas/farmacologia , Feminino , Fibroblastos/efeitos dos fármacos , Fibroblastos/ultraestrutura , Focalização Isoelétrica , Camundongos , Peso Molecular , Ovário , Vibrio/patogenicidade , Virulência
15.
Biochem Cell Biol ; 65(1): 19-26, 1987 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-3566967

RESUMO

The structure of the O-specific polysaccharide of the phenol-soluble cellular lipopolysaccharide of Vibrio anguillarum has been investigated. The studies involved the use of methylation analysis, partial hydrolysis with 48% hydrogen fluoride, Smith degradation, oxidation with chromium trioxide, and comprehensive proton and carbon-13 nuclear magnetic resonance studies, in which one- and two-dimensional experiments were carried out. As a result of these studies it is proposed that the O-specific polysaccharide of Vibrio anguillarum is composed of a regular heteropolymer, i.e., a main chain of (1----4)-linked 3-acetamido-3,6-dideoxy-beta-L-glucose residues alternately substituted through O-2 with side chain residues of 2-acetamido-2,6-dideoxy-alpha-D-glucose, which seem to be substituted through either O-3 or O-4 with propionyl groups (R), as in the following structure. (Formula: see text)


Assuntos
Polissacarídeos Bacterianos , Vibrio/análise , Configuração de Carboidratos , Sequência de Carboidratos , Espectroscopia de Ressonância Magnética , Polissacarídeos Bacterianos/isolamento & purificação
16.
Biokhimiia ; 52(1): 8-14, 1987 Jan.
Artigo em Russo | MEDLINE | ID: mdl-2434145

RESUMO

Electron microscopic analysis of basal bodies of the flagella Vibrio alginolyticus revealed a structure composed of four discs. The diameters of two discs localized in the cytoplasmic membrane appeared to be twice as little as those of the other two discs. In this respect the basal body of V. alginolyticus resembles that of V. cholerae. The 5S sequence of ribosomal RNA from V. alginolyticus appeared to be similar to those of V. cholerae, V. harveyi and some other vibrios. Comparison of 5S-RNA sequence culminated in a dendrogram of evolutionary relationships of various bacterial species, suggesting that V. alginolyticus is a typical representative of the Vibrionacea family. The data obtained are discussed in terms of the role of Na+ energy metabolism in living cells.


Assuntos
Flagelos/ultraestrutura , RNA Bacteriano/análise , RNA Ribossômico/análise , Vibrio/ultraestrutura , Especificidade da Espécie , Vibrio/análise , Vibrio cholerae/análise , Vibrio cholerae/ultraestrutura
17.
J Bacteriol ; 167(1): 57-65, 1986 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-3013839

RESUMO

Anguibactin, a siderophore produced by cells of Vibrio anguillarum 775 harboring the pJM1 plasmid, has now been isolated from the supernatants of iron-deficient cultures. This iron-reactive material was purified by adsorption onto an XAD-7 resin and subsequent gel filtration on a Sephadex LH-20 column. The resulting neutral compound produced an ion at m/z 348 in mass spectrometry and contained one sulfur, four oxygen, and four nitrogen atoms as determined by elemental analysis. Its strong UV absorbance and blue fluorescence were suggestive of a phenolic moiety. In colorimetric reactions anguibactin behaved like a catechol. The catechol assignment was supported by the appearance of a new absorption band at 510 nm in the ferric complex and by the appearance of peaks at 1,367, 1,447, 1,469, and 1,538 cm-1 in the resonance Raman spectrum. In addition, the infrared spectrum gave evidence of a secondary amide function, but no free carboxylic acid or hydroxamic acid groups were observed. A third iron-ligating group was suggested by the liberation of three protons during iron binding; mass spectrometry of the resulting material yielded a molecular ion characteristic of a 1:1 complex of ferric anguibactin. The purified anguibactin exhibited specific growth-promoting activity under iron-limiting conditions for a siderophore-deficient mutant of V. anguillarum 775(pJM1). A novel structure for anguibactin was indicated by the failure of a large number of known siderophores and synthetic chelators to yield a similar type of specific cross-feeding in the V. anguillarum bioassay.


Assuntos
Proteínas da Membrana Bacteriana Externa , Quelantes de Ferro/análise , Peptídeos , Sideróforos , Vibrio/análise , Fenômenos Químicos , Química , Colorimetria , Eletroforese , Quelantes de Ferro/isolamento & purificação , Quelantes de Ferro/metabolismo , Receptores de Superfície Celular/metabolismo , Análise Espectral
18.
J Gen Microbiol ; 132(4): 939-52, 1986 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-3093634

RESUMO

Thermograms obtained by differential scanning calorimetry of a range of bacteria of different heat resistances were compared. Equations were derived to calculate the rate at which the numbers of viable organisms in a calorimeter decline as the temperature is raised at a constant rate. Vegetative bacteria scanned at 10 degrees C min-1 showed multi-peaked thermograms with four major peaks (denoted m, n, p and q) occurring in the regions 68-73, 77-84, 89-99 and 105-110 degrees C respectively. Exceptions were that peak m (the largest peak) occurred at 79-82 degrees C in Bacillus stearothermophilus and an additional peak, r, was detected in Escherichia coli at 119 degrees C. At temperatures below the main peak m there were major differences in thermograms between species. There was a direct relationship between the onset of thermal denaturation and the thermoresistance of different organisms. Heat-sensitive organisms displayed thermogram features which were absent in the more heat-resistant types. When samples were cooled to 5 degrees C and re-heated, a small endothermic peak, pr, was observed at the same temperature as p. Peaks p and pr were identified as the melting endotherms of DNA. In all vegetative organisms examined, maximum death rates, computed from published D and z values, occurred at temperatures above the onset of thermal denaturation, i.e. cell death and irreversible denaturation of cell components occurred within the same temperature range.


Assuntos
Bactérias/análise , Varredura Diferencial de Calorimetria , Calorimetria , Bacillus cereus/análise , Composição de Bases , DNA Bacteriano , Enterococcus faecalis/análise , Escherichia coli/análise , Geobacillus stearothermophilus/análise , Pseudomonas/análise , Streptococcus/análise , Temperatura , Vibrio/análise
19.
Arch Biochem Biophys ; 241(2): 425-31, 1985 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-2994567

RESUMO

A cAMP receptor protein (CRP) species was purified from the luminous Vibrio harveyi cells to apparent homogeneity. This protein had a dimeric structure with a molecular weight of 23,000 per subunit. Among all eight nucleotides tested, only cAMP (Kd = 3 to 4 microM at 0 degrees C and 52 microM at 23 degrees C) and cGMP (Kd = 6 to 10 microM at 0 degrees C and 67 microM at 23 degrees C) bound to this protein. Its binding to poly(dI-dC), poly(dA-dT), and DNA fragments isolated from V. harveyi cells were all significantly enhanced by the addition of cAMP. Based on patterns of limited proteolysis by trypsin, this CRP assumes different conformations in the absence and presence of cAMP. Also consistent with this conclusion is the finding that the binding of cAMP to CRP induced about 50% quenching of the CRP fluorescence with a concomitant 3-nm blue shift from the original 336-nm emission peak. The binding of cGMP resulted in similar fluorescence changes but had no apparent effect on the pattern of proteolysis by trypsin. Using an in vitro transcription system known to be dependent on cAMP and Escherichia coli CRP, the synthesis of a run-off transcript product was also significantly enhanced by cAMP and this V. harveyi CRP.


Assuntos
Receptores de AMP Cíclico/isolamento & purificação , Vibrio/análise , AMP Cíclico/metabolismo , Escherichia coli/análise , Conformação Proteica , Receptores de AMP Cíclico/metabolismo , Transcrição Gênica , Trítio
20.
Infect Immun ; 49(1): 25-31, 1985 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-4008049

RESUMO

Large amounts of an extremely potent extracellular cytolysin produced by the halophilic bacterium Vibrio damsela were obtained free of detectable contamination with medium constituents and other bacterial products by sequential ammonium sulfate precipitation, gel filtration with Sephadex G-100, and hydrophobic interaction chromatography with phenyl-Sepharose CL-4B. The cytolysin is heat labile and protease sensitive and has a molecular weight (estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis) of ca. 69,000 and an isoelectric point of ca. 5.6. The first 10 amino-terminal amino acid residues of the cytolysin are Phe-Thr-Gln-Trp-Gly-Gly-Ser-Gly-Leu-Thr. The cytolysin was very active against erythrocytes from 4 of the 18 animal species examined (mice, rats, rabbits, damselfish) and against Chinese hamster ovary cells and was lethal for mice (ca. 1 microgram/kg, intraperitoneal median lethal dose). Lysis of mouse erythrocytes by the cytolysin is a multi-hit, at least two-step process consisting of a temperature-independent, toxin-binding step followed by a temperature-dependent, membrane-perturbation step(s).


Assuntos
Citotoxinas/isolamento & purificação , Vibrio/análise , Sequência de Aminoácidos , Animais , Citotoxinas/imunologia , Citotoxinas/toxicidade , Hemólise , Temperatura Alta , Imunoeletroforese Bidimensional , Ponto Isoelétrico , Cinética , Camundongos , Peso Molecular
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