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Contribution of the alanine-rich region of Streptococcus mutans P1 to antigenicity, surface expression, and interaction with the proline-rich repeat domain
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Streptococcus mutans is considered to be the major etiologic agent of human dental caries. Attachment of S. mutans to the tooth surface is required for the development of caries and is mediated, in part, by the 185-kDa surface protein variously known as antigen I/II, PAc, and P1. Such proteins are expressed by nearly all species of oral streptococci. Characteristics of P1 include an alanine-rich repeat region and a centrally located proline-rich repeat region. The proline-rich region of P1 has been shown to be important for the translational stability and translocation of P1 through the bacterial membrane. We show here that (i) several anti-P1 monoclonal antibodies require the simultaneous presence of the alanine-rich and proline-rich regions for binding, (ii) the proline-rich region of P1 interacts with the alanine-rich region, (iii) like the proline-rich region, the alanine-rich region is required for the stability and translocation of P1, (iv) both the proline-rich and alanine-rich regions are required for secretion of P1 in Escherichia coli, and (v) in E. coli, P1 is secreted in the absence of SecB. 相似文献
25.
Analysis of the behavior of eryC mutants of Brucella suis attenuated in macrophages 总被引:1,自引:0,他引:1
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The facultatively intracellular pathogen Brucella, characterized by its capacity to replicate in professional and non professional phagocytes, also causes abortion in ruminants. This property has been linked to the presence of erythritol in the placenta, as brucellae preferentially utilize erythritol. The ery operon encodes enzymes involved in erythritol metabolism, and a link with virulence has since been discussed. Allelic exchange mutants in eryC of Brucella suis were erythritol sensitive in vitro with a MIC of 1 to 5 mM of erythritol. Their multiplication in macrophage-like cells was 50- to 90-fold reduced, but complementation of the mutant restored wild-type levels of intracellular multiplication and the capacity to use erythritol as a sole carbon source. In vivo, the eryC mutant colonized the spleens of infected BALB/c mice to a significantly lower extent than the wild type and the complemented strain. Interestingly, eryC mutants that were in addition spontaneously erythritol tolerant nevertheless exhibited wild-type-like intramacrophagic and intramurine replication. We concluded from our results that erythritol was not an essential carbon source for the pathogen in the macrophage host cell but that the inactivation of the eryC gene significantly reduced the intramacrophagic and intramurine fitness of B. suis. 相似文献
26.
Ludwig Wilkens Joelle Tchinda Dagmar Burkhardt Hans Heinrich Kreipe 《Pathobiology》2002,70(4):204-208
OBJECTIVE: Comparative genomic hybridization (CGH) has been established as an informative technique in genetic analysis. However, differences in the ratio of hybridization intensities were reported for particular chromosomes, which may affect CGH results. The aim of this study was to define these differences in more detail. For this purpose, CGH results of 70 samples of bone marrow cells (BMC) with normal karyotype in conventional cytogenetics (CC) were evaluated using seven different reference DNAs and two different DNA labeling systems. METHODS AND RESULTS: CGH using fluorochrome-conjugated nucleotides for DNA labeling indicated signal deviations in 21/70 BMC samples. Deviations affected chromosomes 1 (n = 21), 2 (n = 11), 4 (n = 11), 5 (n = 9), 6 (n = 7), 7 (n = 2), 8 (n = 2), 12 (n = 5), 13 (n = 15), 14 (n = 1), 16 (n = 17), 17 (n = 11), 19 (n = 21), 20 (n = 12), and/or 22 (n = 17). None of the imbalances were confirmed by fluorescence in situ hybridization (FISH). Using digoxigenin and biotin-conjugated nucleotides in exemplary cases (n = 5) led to the disappearance of the signal deviations. Repeated CGH experiments using seven different reference DNAs showed remarkable variations in the signal deviations. CONCLUSION: Hybridization differences depend not only on the hapten or fluorochrome-labeled nucleotides used for DNA labeling, but also on the reference DNA chosen. Therefore, close control of CGH experiments is mandatory, and additional techniques such as FISH should be performed to confirm the results obtained by CGH. 相似文献
27.
Outer membrane protein YadA of enteropathogenic yersiniae mediates specific binding to cellular but not plasma fibronectin. 总被引:7,自引:12,他引:7
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H Schulze-Koops H Burkhardt J Heesemann T Kirsch B Swoboda C Bull S Goodman F Emmrich 《Infection and immunity》1993,61(6):2513-2519
The binding of bacteria or bacterial products to host proteins of tissue extracellular matrix may be a mechanism of tissue adherence. We investigated interactions of the plasmid-encoded outer membrane protein YadA, which confers pathogenic functions on enteropathogenic yersiniae, with fibronectin. Attachment of YadA-positive and YadA-negative recombinant Yersinia enterocolitica strains to cartilage-derived human cellular fibronectin and human plasma fibronectin in the solid phase revealed that YadA mediates binding of yersiniae to cellular fibronectin in a saturable, concentration-dependent manner. The interaction could be inhibited by an anti-YadA-specific anti-serum. An anti-beta 1-integrin antibody and the synthetic peptide G-R-G-D-S-P, representing the binding site for alpha 5 beta 1-integrin on fibronectin, did not block attachment of YadA-positive yersiniae to cellular fibronectin, indicating a binding site for YadA on cellular fibronectin independent of the R-G-D-S-containing site. By contrast, YadA failed to mediate binding to plasma fibronectin immobilized on nitrocellulose or plastic surfaces. These observations provide evidence for the hypothesis that the binding region for YadA in cellular fibronectin is not present in plasma fibronectin. This study is the first report on differential binding of bacteria to splicing variants of fibronectin. Further experiments might answer the question whether binding of YadA to cellular fibronectin contributes to the pathogenesis of yersiniae, both to the initial adhesion of the bacteria to the matrices of the host and to the arthritogenic potential of enteropathogenic yersiniae. 相似文献
28.
Robert Murray John G. Seifert Dennis E. Eddy Gregory L. Paul George A. Halaby 《European journal of applied physiology》1989,59(1-2):152-158
Summary The purpose of this study was to determine the effect of ingesting fluids of varying carbohydrate content upon sensory response,
physiologic function, and exercise performance during 1.25 h of intermittent cycling in a warm environment (T
db=33.4°C). Twelve subjects (7 male, 5 female) completed four separate exercise sessions; each session consisted of three 20
min bouts of cycling at 65%
, with each bout followed by 5 min rest. A timed cycling task (1200 pedal revolutions) completed each exercise session. Immediately
prior to the first 20 min cycling bout and during each rest period, subjects consumed 2.5 ml·kg BW−1 of water placebo (WP), or solutions of 6%, 8%, or 10% sucrose with electrolytes (20 mmol·l−1 Na+, 3.2 mmol·l−1 K+). Beverages were administered in double blind, counterbalanced order. Mean (±SE) times for the 1200 cycling task differed
significantly: WP=13.62±0.33 min, *6%=13.03±0.24 min, 8%=13.30±0.25 min, 10%=13.57±0.22 min (*=different from WP and 10%,P<0.05). Compared to WP, ingestion of the CHO beverages resulted in higher plasma glucose and insulin concentrations, and higher
RER values during the final 20 min of exercise (P<0.05). Markers of physiologic function and sensory perception changed similarly throughout exercise; no differences were
observed among subjects in response to beverage treatments for changes in plasma concentrations of lactate, sodium, potassium,
for changes in plasma volume, plasma osmolality, rectal temperature, heart rate, oxygen uptake, rating of perceived exertion,
or for indices of gastrointestinal distress, perceived thirst, and overall beverage acceptance. Compared to ingestion of a
water placebo, consumption of beverages containing 6% to 10% sucrose resulted in similar physiologic and sensory response,
while ingestion of the 6% sucrose beverage resulted in significantly improved end-exercise performance following only 60 min
of intermittent cycling exercise. 相似文献
29.
H. Hamperl Gsell R. Burkhardt W. Hort Innerhofer F. Schepelmann Wolf H. Braunsteiner J. H. Ellgring R. Gross G. H. Thoenes 《Journal of molecular medicine (Berlin, Germany)》1971,49(23):1301-1304
Ohne Zusammenfassung 相似文献
30.
The ability of currently available anti-arthritic gold preparations to inhibit lysosomal glycosidases from rheumatoid synovial fluid and normal human serum was studied in vitro.It was shown that these preparations differ markedly in their ability to inhibit the enzymes. Gold thioglucose (Solganal) did not inhibit -glucuronidase (-GLUC), -N-acetylglucosaminidase (-NAG) or hyaluronidase (HASE). Chloro(triethylphosphine)gold (SK&F 36914) was a potent inhibitor of -NAG only. Sodium aurothiomalate (Myochrysine and sodium 3-aurothio-2-propanol-1-sulphonate (Allyochrisine) were inhibitors of all three enzymes, notably -GLUC.Kinetic analysis of inhibition by aurothiomalate demonstrated apparent competitive inhibition with -GLUC, but non-competitive inhibition with HASE and -NAG -GLUC was also strongly inhibited by silver and copper thiomalates.The concentrations of these drugs required for effective inhibition of lysosomal glycosidases probably exceed those attained in serum and therefore preclude this action extracellularly. It is suggested that durg sequestration and retention within phagocytic cells facilitates inhibition of glycosaminoglycan catabolism that mediates cleavage of glucuronidic linkages of hyaluronic acid and chondroitin sulphates.The hypothesis that gold dompounds act in vivo by attenuating the activity of lysosomal enzymes is discussed in relation to these and previous findings. 相似文献