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61.
62.
Fulde M Rohde M Hitzmann A Preissner KT Nitsche-Schmitz DP Nerlich A Chhatwal GS Bergmann S 《The Biochemical journal》2011,434(3):523-535
Streptococcus canis is an important zoonotic pathogen capable of causing serious invasive diseases in domestic animals and humans. In the present paper we report the binding of human plasminogen to S. canis and the recruitment of proteolytically active plasmin on its surface. The binding receptor for plasminogen was identified as a novel M-like protein designated SCM (S. canis M-like protein). SPR (surface plasmon resonance) analyses, radioactive dot-blot analyses and heterologous expression on the surface of Streptococcus gordonii confirmed the plasminogen-binding capability of SCM. The binding domain was located within the N-terminus of SCM, which specifically bound to the C-terminal part of plasminogen (mini-plasminogen) comprising kringle domain 5 and the catalytic domain. In the presence of urokinase, SCM mediated plasminogen activation on the bacterial surface that was inhibited by serine protease inhibitors and lysine amino acid analogues. Surface-bound plasmin effectively degraded purified fibrinogen as well as fibrin clots, resulting in the dissolution of fibrin thrombi. Electron microscopic illustration and time-lapse imaging demonstrated bacterial transmigration through fibrinous thrombi. The present study has led, for the first time, to the identification of SCM as a novel receptor for (mini)-plasminogen mediating the fibrinolytic activity of S. canis. 相似文献
63.
Conserved sequence motifs, alignment, and secondary structure for the third domain of animal 12S rRNA 总被引:17,自引:3,他引:14
Hickson RE; Simon C; Cooper A; Spicer GS; Sullivan J; Penny D 《Molecular biology and evolution》1996,13(1):150-169
Secondary structure models are an important step for aligning sequences,
understanding probabilities of nucleotide substitutions, and evaluating the
reliability of phylogenetic reconstructions. A set of conserved sequence
motifs is derived from comparative sequence analysis of 184 invertebrate
and vertebrate taxa (including many taxa from the same genera, families,
and orders) with reference to a secondary structure model for domain III of
animal mitochondrial small subunit (12S) ribosomal RNA. A template is
presented to assist with secondary structure drawing. Our model is similar
to previous models but is more specific to mitochondrial DNA, fitting both
invertebrate and vertebrate groups, including taxa with markedly different
nucleotide compositions. The second half of the domain III sequence can be
difficult to align precisely, even when secondary structure information is
considered. This is especially true for comparisons of anciently diverged
taxa, but well-conserved motifs assist in determining biologically
meaningful alignments. Patterns of conservation and variability in both
paired and unpaired regions make differential phylogenetic weighting in
terms of "stems" and "loops" unsatisfactory. We emphasize looking carefully
at the sequence data before and during analyses, and advocate the use of
conserved motifs and other secondary structure information for assessing
sequencing fidelity.
相似文献
64.
All 25 cultures of Actinomyces pyogenes tested in the present study bound 125I-labelled human alpha 2-macroglobulin with a mean binding of 65.6%. Thirteen cultures also bound 125I-labelled human haptoglobin with a mean of 51.5%. None interacted with fibrinogen, fibronectin, immunoglobulin G, or albumin. Twenty-eight cultures representing other species of actinomycetaceae did not show any interaction with alpha 2-macroglobulin, haptoglobin, and other plasma proteins tested. The binding of alpha 2-macroglobulin and haptoglobin to A. pyogenes was saturable and could be completely inhibited by the respective unlabelled plasma proteins. The binding of alpha 2-macroglobulin could not be inhibited by unlabelled haptoglobin. On the other hand, alpha 2-macroglobulin blocked the binding of haptoglobin, possibly by steric hindrance. Treatment of the bacteria with trypsin reduced their binding activities for alpha 2-macroglobulin and haptoglobin indicating the protein nature of the binding sites. Exposure to heat (1 h, 80 degrees C) significantly diminished the binding activity for haptoglobin, but not that for alpha 2-macroglobulin. The binding of alpha 2-macroglobulin and haptoglobin could be an important feature in the classification of A. pyogenes among the members of actinomycetaceae. 相似文献
65.
A simple method for quantitative determination of bacterial adherence to human and animal epithelial cells 总被引:3,自引:0,他引:3
A simple quantitative method for determining bacterial adherence to epithelial cells was devised. The method involved incubation of fluorescein-labeled bacteria with oral epithelial cells. Non-adherent bacteria were subsequently removed by Percoll density centrifugation. The degree of bacterial adherence was then determined fluorospectrophotometrically by measuring the fluorescein extracted from the adherent bacteria. The degree of adherence corresponded well with the microscopic counts of adherent bacteria. The method proved to be suitable for assaying adherence of both gram-positive and gram-negative bacteria to human as well as animal epithelial cells. 相似文献
66.
Synthesis and immunomodulating tumor control properties of albumin covalently connected to trehalose
Walter J. Gensler Vijay Kumar Chhatwal Iftekhar Alam Edwin Constantino George S. Tarnowski M. V. Pimm R. W. Baldwin 《Cancer immunology, immunotherapy : CII》1980,9(1-2):101-109
Summary Proteins conjugated to a trehalose derivative are described. These include human serum albumin and bovine serum albumin covalently attached through appropriate molecular linkages to the 6- and 6,6-positions of trehalose. The immunomodulating tumor control activities of these conjugated proteins has been assayed in several test systems. 相似文献
67.
Abstract A multidisciplinary field study investigating the fate and transport of petroleum hydrocarbons commonly associated with jet-fuel contamination is currently underway at Columbus Air Force Base (AFB), Mississippi. Sixty sediment cores from 12 boreholes were recovered from the study aquifer. The goal of this initial sampling was to characterize the potential microbial activity using 14C-labeled substrates, as well as the presence, abundance, and distribution of specific hydrocarbon degrading genotypes using DNA:DNA hybridization. Enumeration of total microbial abundance using a 16S rDNA universal oligonucleotide probe was compared to traditional enumeration methods. Total culturable populations determined by spread plate analysis ranged from a low of 10(4) to more than 10(6) organisms per gram sediment. Microbial abundance estimated by DNA hybridization studies with 16S rDNA genes ranged from 10(7) to 10(8) organisms per gram sediment. Molecular analysis of aquifer samples using DNA probes targeting genes encoding the degradative enzymes alkane hydroxylase (alkB), catechol 2,3-dioxygenase (nahH), naphthalene dioxygenase (nahA), toluene dioxygenase (todC1C2), toluene monooxygenase (tomA), and xylene monooxygenase (xylA), as well as two probes measuring methanogenic microorganisms, codh (carbon monoxide dehydrogenase) and mcr (methyl coenzyme reductase), revealed that each target gene sequence was present in nearly all 60 samples. The presence of organisms demonstrating the phenotype to degrade BTEX and naphthalene was further supported using mineralization assays with 14C-labeled benzene, toluene, naphthalene, and phenanthrene. Minimal activity occurred during the first 24 hours. After a period of 5-7 days, greater than 40% of the target compounds were mineralized in aquifer sediments. 相似文献
68.
Domain structure and conserved epitopes of Sfb protein, the fibronectin-binding adhesin of Streptococcus pyogenes 总被引:9,自引:3,他引:6
Susanne R. Talay Peter Valentin-Weigand Kenneth N. Timmis Gursharan S. Chhatwal 《Molecular microbiology》1994,13(3):531-539
Streptococcus pyogenes expresses a fibronectin-binding surface protein (Sfb protein) which mediates adherence to human epithelial cells. The nucleotide sequence of the sfb gene was determined and the primary sequence of the Sfb protein was analysed. The protein consists of 638 amino acids and comprises five structurally distinct domains. The protein starts with an N-terminal signal peptide followed by an aromatic domain. The central part of the protein is formed by four proline-rich repeats which are flanked by non-repetitive spacer sequences. A second repeat region, consisting of four repeats that are distinct from the proline repeats and have been shown to form the fibronectin-binding domain, is located in the Cterminal part of the protein. The protein ends with a typical cell wall and membrane anchor region. Comparative sequence analysis of the N-terminal aromatic domain revealed similarities with carbohydrate-binding sites of other proteins. The proline repeat region of the Sfb protein shares characteristic features with proline-rich repeats of functionally distinct surface proteins from pathogenic Gram-positive cocci. Immunoelectron microscopy revealed an even distribution of the fibronectin-binding domain of Sfb protein on the surface of streptococcal cells. Analyses of 38 sfb genes originating from different S. pyogenes isolates revealed primary sequence variability in regions coding for the N-termini of mature Sfb proteins, whereas sequences coding for the central and C-terminal repeats were highly conserved. The repeat sequences are postulated to act as target sites for intragenic recombination events that result in variable numbers of repeats within the different sfb genes. A model of the Sfb protein is presented. 相似文献
69.
Aflatoxin G1 (AFG1) was transformed into aflatoxin B3 (AFB3) by the fungiRhodotorula sp,Sporobolomyces sp,Rhizopus oryzae NRRL395,Pythium ultimum, Aspergillus terreus, A clavatus and Penicillium frequentans grown in a medium containing AFG1, Difco potato dextrose broth, yeast extract, and peptone both in liquid shaken cultures and in solid static cultures at 25°C in the dark. A maximum rate of transformation of 10 % was obtained after 2 to 3 weeks of incubation. The transformation was correlated with an increase in the pH of the media from 5.7 –5.9 to 8.3 – 8.8.Saccharomyces cerevisiae also transformed AFG1 into AFB3, but at a slower rate; the pH of the media did not reach above 8.0 until 5 weeks after incubation. No transformation was observed whenA niger andP chrysogenum were tested; in both cases, no increase in pH was noticed. However, some transformation of AFG1 to AFB3 by both fungi was observed when the initial pH of the media was adjusted to 9.0. The rate of transformation increased to 15 – 20% in the static culture where the same medium was adsorbed onto vermiculite andRhizopus andAspergilli gave the highest increase in AFB3 yield. 相似文献
70.