共查询到20条相似文献,搜索用时 15 毫秒
1.
A larger-molecular-weight precursor of enterotoxin A was found in membranes of Staphylococcus aureus and was shown to be the kinetic precursor to the extracellular form of the toxin. Subcellular fractionation revealed that mature enterotoxin A was transiently associated with the cell wall before being released to the extracellular environment. 相似文献
2.
A larger, membrane-bound form of staphylococcal enterotoxin B was shown by in vivo pulse-chase analysis to be the kinetic precursor to extracellular enterotoxin B. Processing of the enterotoxin B precursor molecules can apparently occur either cotranslationally or posttranslationally. Subcellular fractionation of cells revealed that all of the precursor toxin was associated with the membrane fraction. Once processed and released from the membrane, it was transiently associated with the cell wall before being released into the extracellular environment. The cell-wall-associated enterotoxin B was completely resistant to protease treatment and to extraction by high- or low-salt solutions at 0 to 2 degrees C, although it could be easily released from the cell by removal of the cell wall with lysostaphin. These data imply that newly formed enterotoxin B may be temporarily sequestered in specialized regions that require cell wall integrity before being released into the extracellular environment. 相似文献
3.
Nucleotide sequence of the type C3 staphylococcal enterotoxin gene suggests that intergenic recombination causes antigenic variation. 总被引:12,自引:1,他引:11
下载免费PDF全文
![点击此处可从《Journal of bacteriology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
The nucleotide sequence of the structural gene for staphylococcal enterotoxin type C3 (entC3) was determined. This gene contains 798-base-pair open reading frame that encodes a protein of 266 amino acid residues. Sequence analysis suggests that staphylococcal enterotoxin type C3 is synthesized in a precursor form that is processed to yield a mature extracellular form of 238 amino acid residues (molecular weight, 27,438). The entC3 gene is closely related to the gene for staphylococcal enterotoxin type C1, with 98% nucleotide sequence identity. Sequence comparisons between the entC3, entC1, and entB genes suggest that an ancestral entC1-like gene was formed by recombination between the entC3 and entB genes. 相似文献
4.
Clostridium perfringens type A enterotoxin (CPE) has been shown previously to inhibit the incorporation of radiolabeled precursors into acid-insoluble material but the mechanism of inhibition is unknown. It has also been shown that extracellular calcium is required for some CPE effects. In this report, it is shown that CPE completely and virtually simultaneously inhibits incorporation of precursors into RNA, DNA and protein in either the presence or absence of extracellular divalent cations and that changes in intracellular precursor levels did not consistently correlate with this CPE-induced inhibition of incorporation. These results strongly suggest that CPE can inhibit macromolecular synthesis, not just inhibit precursor transport. It is inferred from this that CPE can affect DNA and RNA synthesis, and possibly protein synthesis, by altering other cellular processes besides, or in addition to, precursor transport and these effects then lead to a shutdown of macromolecular synthesis. 相似文献
5.
Cell associated staphylococcal enterotoxin A, released by lysostaphin treatment of Staphylococcus aureus cells, was found to be biologically active in cynomolgus monkeys. The activity was comparable to that of extracellular enterotoxin A; six of six monkeys vomited within 5 h in response to extracellular enterotoxin and four of six also vomited in response to the same serologic level (4.8 μg per monkey) of cell-associated enterotoxin. Feeding S. aureus cells containing cell-associated enterotoxin A to cynomolgus monkeys resulted in emesis in three of five monkeys within 3 h. This suggests that consumption of S. aureus cells could lead to staphylococcal intoxication. 相似文献
6.
大肠杆菌肠毒素的基因融合及其免疫原性的研究 总被引:4,自引:0,他引:4
应用重组基因工程技术,将热敏肠毒素B亚基(LT-B)基因与含有部分前体的耐热肠毒素(pro-ST)基因融合在一起,构建了表达LT-B/pro-ST融合蛋白的重组质粒.该融合蛋白不仅保持结合神经节苷脂(GM1)的能力,而且具有热敏肠毒素和耐热肠毒素的抗原性.乳鼠实验证明,融合蛋白虽然含有野生耐热肠毒素,但不具耐热肠毒素的生物毒性.腹腔免疫和鼻饲免疫均能激发产生抗ETEC两种肠毒素的抗体. 相似文献
7.
Vibrio cholerae enterotoxin genes: nucleotide sequence analysis of DNA encoding ADP-ribosyltransferase. 总被引:8,自引:2,他引:6
下载免费PDF全文
![点击此处可从《Journal of bacteriology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
The nucleotide sequence of the DNA encoding the ADP-ribosyltransferase (A1) fragment of cholera enterotoxin was determined. A putative precursor of the A1 peptide contains an 18-amino acid leader peptide, and the mature A1 peptide contains 194 amino acids. The primary structure of the A1 fragment from cholera enterotoxin is more related to that from a human enterotoxigenic Escherichia coli than to that from a porcine enterotoxigenic E. coli. 相似文献
8.
Synthesis of Escherichia coli heat-stable enterotoxin STp as a pre-pro form and role of the pro sequence in secretion. 总被引:9,自引:3,他引:6
下载免费PDF全文
![点击此处可从《Journal of bacteriology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
Escherichia coli heat-stable enterotoxin STp is presumed from its DNA sequence to be synthesized in vivo as a 72-amino-acid residue precursor that is cleaved to generate mature STp consisting of the 18 carboxy-terminal amino acid residues. There are two methionine residues in the inferred STp sequence in addition to the methionine residue at position 1. In order to confirm production of the STp 72-amino-acid residue precursor, we substituted the additional methionine residues by oligonucleotide-directed site-specific mutagenesis. Since these substitutions did not cause a significant change in STp production, it can be concluded that STp is normally synthesized as the 72-amino-acid residue precursor. The length of the STp precursor indicated the existence of a pro sequence between the signal peptide and the mature protein. In order to identify the pro sequence and determine its role in protein secretion, deletion and fusion proteins were made. A deletion mutant in which the gene fragment encoding amino acid residues 22 to 53 of STp was removed was made. STp activity was found in the culture supernatant of cells. Amino acid sequence analysis of the purified STp deletion mutant revealed that the pro sequence encompasses amino acid residues 20 to 54. A hybrid protein consisting of STp amino acids 1 to 53 fused in frame from residue 53 to nuclease A was not secreted into the culture supernatant. These results indicate that the pro sequence does not function to guide periplasmic protein into the extracellular milieu. 相似文献
9.
10.
The production of enterotoxins, lipase and total extracellular protein by four strains of Staphylococcus aureus grown in batch culture at a controlled pH of 6.5 in a completely defined medium was markedly reduced by glucose or glycerol constantly maintained at 0.I M. A concomitant increase in the production of deoxyribonuclease, up to 13-fold, showed however that not all extracellular proteins are under the same control mechanism. The presence of glucose and glycerol in the medium also resulted in a rapid increase in the specific growth rate. However, growth of S. aureus s6 in Mgilimited continuous culture showed that glucose repression of enterotoxin B when the growth rate was held constant was more than twice that in batch culture. Therefore glucose repression can occur independently of an increase in growth rate. The specific rate of production of enterotoxin B, lipase, deoxyribonuclease, beta-haemolysin and total extracellular protein by S. aureus s6 increased as the growth rate increased from 0.07 to 0.24 h-1. Non-replicating cells grown in the absence of glucose produced considerable amounts of enterotoxin, and production was not repressed by the presence of glucose in the resuspension medium. In contrast, no enterotoxin B or C was obtained from nonreplicating cells grown in the presence of glucose. Chloramphenicol completely inhibited enterotoxin production by non-replicating cells, indicating that synthesis of new protein was required. 相似文献
11.
Cloning and nucleotide sequence of the type E staphylococcal enterotoxin gene. 总被引:34,自引:5,他引:29
下载免费PDF全文
![点击此处可从《Journal of bacteriology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
The gene for staphylococcal enterotoxin type E (entE) was cloned from Staphylococcus aureus into plasmid vector pBR322 and introduced into Escherichia coli. A staphylococcal enterotoxin type E-producing E. coli strain was isolated. The complete nucleotide sequence of the cloned structural entE gene and the N-terminal amino acid sequence of mature staphylococcal enterotoxin type E were determined. The entE gene contained 771 base pairs that encoded a protein with a molecular weight of 29,358 which was apparently processed to a mature extracellular form with a molecular weight of 26,425. DNA sequence comparisons indicated that staphylococcal enterotoxins type E and A are closely related. There was 84% nucleotide sequence homology between entE and the gene for staphylococcal enterotoxin type A; these genes encoded protein products that had 214 (83%) homologous amino acid residues (mature forms had 188 [82%] homologous amino acid residues). 相似文献
12.
Secretion of methanol-insoluble heat-stable enterotoxin (STB): energy- and secA-dependent conversion of pre-STB to an intermediate indistinguishable from the extracellular toxin. 总被引:13,自引:2,他引:11
下载免费PDF全文
![点击此处可从《Journal of bacteriology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
Y M Kupersztoch K Tachias C R Moomaw L A Dreyfus R Urban C Slaughter S Whipp 《Journal of bacteriology》1990,172(5):2427-2432
The methanol-insoluble, heat-stable enterotoxin of Escherichia coli synthesized by clinical strains or strains that harbor the cloned gene was shown to be an extracellular polypeptide. The toxin (STB) was first detected as an 8,100-Mr precursor (pre-STB) that was converted to a transiently cell-associated 5,200-Mr form. Proteolytic conversion of pre-STB to STB was shown to be inhibited by the proton motive force uncoupler carbonyl cyanide m-chlorophenylhydrazone and did not occur in a secA background. After STB was detected as a cell-associated molecule, an extracellular form with identical electrophoretic mobility became apparent. The results suggest that there is no proteolytic processing during the mobilization of STB from the periplasm to the culture supernatant. The determined amino acid sequence of STB coincides fully with the 48 carboxy-terminal amino acids inferred from the DNA sequence. The 23 amino-terminal residues inferred from the DNA sequence were absent in the mature toxin. 相似文献
13.
Hasegawa M Kawano Y Matsumoto Y Hidaka Y Fujii J Taniguchi N Wada A Hirayama T Shimonishi Y 《Protein expression and purification》1999,15(3):271-281
Guanylyl cyclase (GC)-C, a single-transmembrane receptor protein for heat-stable enterotoxin, guanylin, and uroguanylin, and its N-terminal extracellular domain were prepared at a high level of expression from a system constructed of Sf21 insect cells and recombinant baculovirus. The recombinant GC-C, containing the complete sequence, retained its binding affinity to heat-stable enterotoxin with a KD value (6.2 x 10(-10) M) and cyclase catalytic activity at a level similar to those of GC-C expressed in mammalian cell lines, such as COS-7. The N-terminal extracellular domain was prepared in a form which contained the hexahistidine tail at its C-terminus and was purified as a homogenous protein by Con A and Ni-chelating affinity chromatography from the culture medium of the insect cells. The purified N-terminal extracellular domain of GC-C exhibited the high (KD = 4 x 10(-10) M) and low (KD = 7 x 10(-8) M) affinity sites in binding to heat-stable enterotoxin. These results clearly indicate that the N-terminal extracellular domain of GC-C possesses the same biochemical characteristics as the complete GC-C protein even in the membrane-free form. Moreover, the extracellular domain is able to form an oligomer in a ligand-dependent manner, suggesting that the N-terminal extracellular domains interact with one another in binding to ligands. 相似文献
14.
15.
Production of Escherichia coli Heat-labile Enterotoxin in Fermenter Dialysis Culture 总被引:1,自引:1,他引:0
Production of Escherichia coli heat-labile enterotoxin was investigated with one porcine and one human strain in three different media under different cultivation conditions. Cultivation in aerated fermenters at pH 7·0 yielded 10–20 times more enterotoxin/ml of culture fluid than cultivation in shake flasks. A trypton-yeast extract medium was optimal in fermenter cultures. Comparatively good yields of enterotoxin in fermenters were also obtained in a glucose-salts medium. Continuous feeding of glucose and salts during fermenter cultivation resulted in a lower production of enterotoxin/mg of bacterial cells. Since this decrease in specific yield could be reversed by using dialysis culture, it was concluded that inhibition of toxin formation was due to the accumulation of extracellular low molecular weight metabolites. The highest yield of enterotoxin in dialysis culture was 80 ED50 ml−1 (rabbit jejunal loop test) which is at least eight times more toxin than in ordinary fermenter culture and 80 times more toxin than in shake flask cultures. 相似文献
16.
Nucleotide sequence of the staphylococcal enterotoxin C3 gene sequence comparison of all three type C staphylococcal enterotoxins 总被引:14,自引:0,他引:14
Carolyn J. Hovde Sean P. Hackett Gregory A. Bohach 《Molecular & general genetics : MGG》1990,220(2):329-333
Summary The structural gene entC3, which encodes staphylococcal enterotoxin C3 was cloned from the genome of Staphylococcus aureus FRI-913 and sequenced. The primary amino acid sequence of the toxin was deduced from the nucleotide sequence data. entC3 contains 801 by and encodes a precursor protein of 266 amino acids. Glutamic acid was found to be the N-terminus of mature enterotoxin C3. Thus, the first 27 residues of the toxin precursor comprise the signal peptide, and the mature toxin contains 239 amino acids with a molecular weight of 27 563 daltons. Enterotoxin C3 differs from enterotoxin C2 by four amino acids and from enterotoxin C1 by nine residues. The 167 C-terminal residues of the three toxins are identical, except for one conservative amino acid substitution in enterotoxin C3. The degree of immunological relatedness among the three Type C enterotoxins is proportional to their molecular relatedness. This study also provides evidence that the N-termini of Type C enterotoxins determine subtype-specific antigenic epitopes, while more conserved C-terminal regions determine biological properties and cross-reactive antigenic epitopes shared with other pyrogenic toxins. 相似文献
17.
Transient entry of enterotoxin subunits into the periplasm occurs during their secretion from Vibrio cholerae. 总被引:26,自引:9,他引:17
下载免费PDF全文
![点击此处可从《Journal of bacteriology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
Cholera toxin and heat-labile enterotoxin (LT) are structurally similar oligomeric proteins which are capable of being efficiently secreted from Vibrio cholerae. Here we report that these proteins transiently enter the periplasm of V. cholerae as they traverse the cell envelope to reach the extracellular milieu. Pulse-chase experiments on V. cholerae TRH7000 harboring an LT-encoding plasmid revealed that radiolabeled LT A and B subunits entered the periplasm rapidly, followed by their slow efflux (half-time, 13 min) into the medium. LT B-subunit efflux from the periplasm was calculated to be at a rate of ca. 170 monomers per min per cell (which is equivalent to 34 assembled LT holotoxin molecules per min per cell). These values were estimated to be sufficient to account for the increase in extracellular enterotoxin concentration during exponential cell growth. Thus, all enterotoxin subunits which are secreted into the medium can be assumed to be channelled via the periplasm. These findings led to an improved model of the pathway of toxin secretion by V. cholerae. 相似文献
18.
We determined the nucleotide sequence of the gene encoding staphylococcal enterotoxin A (entA). The gene, composed of 771 base pairs, encodes an enterotoxin A precursor of 257 amino acid residues. A 24-residue N-terminal hydrophobic leader sequence is apparently processed, yielding the mature form of staphylococcal enterotoxin A (Mr, 27,100). Mature enterotoxin A has 82, 72, 74, and 34 amino acid residues in common with staphylococcal enterotoxins B and C1, type A streptococcal exotoxin, and toxic shock syndrome toxin 1, respectively. This level of homology was determined to be significant based on the results of computer analysis and biological considerations. DNA sequence homology between the entA gene and genes encoding other types of staphylococcal enterotoxins was examined by DNA-DNA hybridization analysis with probes derived from the entA gene. A 624-base-pair DNA probe that represented an internal fragment of the entA gene hybridized well to DNA isolated from EntE+ strains and some EntA+ strains. In contrast, a 17-base oligonucleotide probe that encoded a peptide conserved among staphylococcal enterotoxins A, B, and C1 hybridized well to DNA isolated from EntA+, EntB+, EntC1+, and EntD+ strains. These hybridization results indicate that considerable sequence divergence has occurred within this family of exotoxins. 相似文献
19.
Regulation of the enterotoxin B gene in Staphylococcus aureus 总被引:12,自引:0,他引:12
20.
Thermoactivation of a periplasmic heat-stable enterotoxin of Escherichia coli. 总被引:7,自引:4,他引:3
下载免费PDF全文
![点击此处可从《Journal of bacteriology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
Strains of Escherichia coli that host a plasmid that codes for the heat-stable (ST) enterotoxin showed 160 times more extracellular enterotoxin than intracellular activity. However, when washed bacteria were sonicated and incubated at between 50 and 85 degrees C, an activity similar to that of the ST enterotoxin was detected. No such effect was present in strains lacking the plasmid, in a plasmid ST- mutant, or in chromosomal mutants that lack a cyclic AMP-linked positive regulatory system which previously were shown to yield an ST- phenotype. The thermoactivation was inhibited by iodoacetamide and N-ethylmaleimide; chloramphenicol did not affect the phenomenon. The heat-activated ST-like enterotoxin was localized in the periplasmic space. The results are discussed in relation to the export of the toxin from the periplasm to the outside of the cell. 相似文献