Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

---

Query: EC:3.4.21.4 (trypsin)
42,187 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

We have recently reported that three distinct size- and phase-variable surface lipoproteins (Vsps) of the bovine pathogen Mycoplasma bovis possess a common epitope recognized by monoclonal antibody 1E5. In the present study, we show that this epitope is also present on a size-variant protein (PvpA) of the avian pathogen Mycoplasma gallisepticum. Application of monoclonal antibody 1E5 in Western immunoblot analysis of Triton X-114 phase-fractionated proteins and in colony immunoblots, as well as in trypsin and carboxypeptidase digestion experiments, has demonstrated that (i) PvpA is an integral membrane protein with a free C terminus, (ii) the shared epitope is surface exposed, and (iii) PvpA is subjected to high-frequency phase variation in expression. By using serum antibodies from M. gallisepticum-infected chickens, we were able to demonstrate the immunogenic nature of PvpA and identify three additional highly immunogenic Triton X-114 phase proteins (p67, p72, and p75) also undergoing high-frequency phase variation spontaneously and independently. Metabolic labeling experiments with [14C]palmitate and [14C]oleate revealed that PvpA, in contrast to p67, p72, and p75, is not lipid modified. Southern blot hybridization with restriction fragments carrying the pvpA gene of M. gallisepticum or the vspA gene of M. bovis against digested genomic DNA of the two Mycoplasma species indicated the absence of genetic relatedness between the pvpA and vspA genes. The apparent complexity of the antigenic variation phenomenon in M. gallisepticum is discussed.
...
PMID:A surface epitope undergoing high-frequency phase variation is shared by Mycoplasma gallisepticum and Mycoplasma bovis. 752 2

An antigen containing a common epitope in most strains of Mycoplasma gallisepticum was purified by isoelectric focusing and used in the production of monoclonal antibodies (mAb). Of several mAb produced, only one mAb reacted with focused component and with all six strains of M. gallisepticum except strain 6/85. This mAb was designated MG3D6.A5, and it was subsequently purified with immobilized rProtein Atm. The MG3D6.A5 mAb recognized a common epitope on a molecule with relative molecular weight of 98 kilodaltons (kDa), termed p98. No binding was observed when the MG3D6.A5 mAb was reacted against antigens extracted from other mycoplasma species, indicating its species-specificity. Physicochemical studies revealed that p98 had an isoelectric point of 5.2, was stable to heat, and was resistant to periodate oxidation but sensitive to trypsin treatment, suggesting that p98 is a nonglycosylated protein. Furthermore, ultrastructural studies with colloidal gold revealed that M. gallisepticum cells were selectively stained with MG3D6.A5 mAb to p98. The latter was focally distributed on the surface of a mycoplasma cell membrane near the attachment organelle. These results suggest that p98 is a highly conserved protein in M. gallisepticum strains, is immunogenic, and is surface-accessible; its binding specificity to MG3D6.A5 mAb could be used to identify M. gallisepticum in multiple cultures.
...
PMID:Evidence for a common epitope on the surface of Mycoplasma gallisepticum defined by monoclonal antibody. 768 75

Human immunoglobulin G Fc fragment-binding activity of Mycoplasma salivarium cells was remarkably enhanced by trypsin treatment of the cells. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis profile of proteins of the cells treated with trypsin was the same as that of the cells treated with pronase, although pronase treatment had been shown to reduce the activity in our previous study (FEMS Microbiol. Lett. 123, 305-310, 1994). This contradiction was clarified by the finding that trypsin bound the Fc fragment more strongly than the cells, and a small amount of trypsin remained in the cells treated with trypsin and washed well. On the basis of these results, it was concluded that the enhancement of cell activity by trypsin treatment was ascribed to binding of the Fc fragment to trypsin remaining in the trypsin-treated cells.
...
PMID:The enhancement of the human immunoglobulin G Fc fragment-binding activity of Mycoplasma salivarium cells by trypsin treatment is ascribed to the binding of trypsin to the Fc fragment. 774 43

Results of competitive binding studies with radiolabeled and unlabeled Mycoplasma pulmonis and rat tracheal explant cultures indicated no effect of trypsin treatment on the ability of M. pulmonis to bind to explants. A trypsin-resistant 46-kDa membrane protein, which binds isolated rat tracheal epithelial cells in culture, was purified and used to produce specific antibodies that block adhesion of mycoplasmas to tracheal explants. These results suggest that M. pulmonis adheres to rat tracheal epithelium via a trypsin-resistant 46-kDa protein.
...
PMID:Mycoplasma pulmonis 46-kDa trypsin-resistant protein adheres to rat tracheal epithelial cells. 789 30

The protein and antigen profiles of 11 isolates of Mycoplasma bovis were compared by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblot analysis of whole organisms. The isolates examined included the type strain PG45 and 10 other filter-cloned strains or purified isolates both from animals without clinical signs and from clinical cases of bovine mastitis, arthritis, or pneumonia. While the overall protein patterns visualized by silver staining were very similar, marked differences in the antigen banding profiles were detected by rabbit antiserum prepared against whole organisms from one of the strains analyzed. This antigenic heterogeneity was shown to be independent of the geographical origin, the type of clinical disease, and the site of isolation and was also observed among serial isolates from a single animal. Antigen profiles were further monitored throughout sequentially subcloned populations of the PG45 strain. This clonal analysis revealed a high-frequency variation in the expression levels of several prominent antigens. All of these variable antigens were defined by detergent-phase fractionation with Triton X-114 as amphiphilic integral membrane proteins. A subset of different-sized membrane proteins was identified by a monoclonal antibody raised against a PG45 subclone expressing a 63- and a 46-kDa variant antigen within that set. The selective susceptibility of these proteins to trypsin treatment of intact organisms and their ability to bind the monoclonal antibody in colony immunoblots demonstrated that they were exposed on the cell surface. In addition, their preferential recognition by serum antibodies from individual cattle with naturally induced M. bovis mastitis or arthritis confirmed that they were major immunogens of this organism. These studies establish that the apparent antigenic heterogeneity among M. bovis isolates reported here does not represent stable phenotypic strain differences generated from accumulated mutational events but reflects distinct expression patterns of diverse, highly variable membrane surface proteins.
...
PMID:Antigen heterogeneity among isolates of Mycoplasma bovis is generated by high-frequency variation of diverse membrane surface proteins. 792 89

A highly hydrophobic component derived from the membrane of Mycoplasma capricolum has been characterized, purified and assessed for its ability to activate macrophages to tumor cytotoxicity. Initially, crude membranes were evaluated for their solubility in a wide range of solvents. Despite differential solubility in the various solvents, the mycoplasma membranes retained their ability to potentiate macrophage tumor cytotoxicity. Mycoplasma membranes were further characterized by appraising their macrophage-activating ability subsequent to various chemical treatments: cleavage of ester and thioester bonds, oxidation of vicinal hydroxyl groups, and exposure to a broad range of pH. Only strong alkaline treatment (pH > 12) caused a reduction in mycoplasma membrane activity; all other chemical treatments were inconsequential. With potential therapeutic applications in mind, mycoplasma membranes were subjected to various physical treatments including heating, freezing/thawing, sonication, lyophilization and storage. The ability of the membranes to induce macrophage activation was stably maintained following all these treatments. Purification of membranes was initiated by a chloroform/methanol lipid extraction. Macrophage-activating ability was found predominantly in the interphase. Proteolytic cleavage with trypsin increased specific activity at least sixfold. Trypsinized fractions were solubilized in 2-chloroethanol and gel filtration was performed on a hydroxylated Sephadex LH-60 column. The active fraction from this column had a further tenfold increase in specific activity. Subsequent rounds of reverse-phase HPLC on this fraction yielded three to four peaks absorbing at 280 nm, of which only one had macrophage-activating ability.
...
PMID:Characterization and purification of a mycoplasma membrane-derived macrophage-activating factor. 804 23

Adherence of Mycoplasma hyopneumoniae to the mucosa of the distal portion of the respiratory tract of swine is an important initial event in development of mycoplasmal pneumonia. A suitable in vitro model of adherence would be useful for investigation of mycoplasmal and host cell factors involved in this process. We have developed an adherence assay, using suspensions of porcine respiratory tract ciliated epithelial cells and M hyopneumoniae. Tracheal epithelial cells, collected by use of cytologic brushes, were mixed with broth cultures of M hyopneumoniae and the mixtures were incubated, diluted, vortexed, and sedimented. Pellets were spread on glass slides, stained with a fluorescent antibody against M hyopneumoniae, and evaluated by fluorescent microscopy. Fluorescence was observed principally among cilia on the ciliated tufts of epithelial cells. Only a few organisms were observed adhering on the nonciliated parts of ciliated cells or on other cell types. When mycoplasmas were preincubated with low dilutions of serum from swine convalescing from M hyopneumoniae disease, attachment was partially inhibited (P < 0.05). Significant inhibition of attachment was not observed when organisms were preincubated with higher dilutions of convalescent serum, with purified IgG from hyperimmune serum against M hyopneumoniae, or with low dilutions of lung lavage fluids (from convalescent swine) that contained specific IgA antibodies against M hyopneumoniae. Preincubation of the organisms with periodate and trypsin abolished attachment and formaldehyde decreased it (P < 0.05), whereas a variety of carbohydrates had no effect on attachment. Preincubation with dextran sulfate, ammonium sulfate, magnesium sulfate, and methionine reduced attachment (P < 0.05). Treatment of cell-Mycoplasma mixtures with the hydrophobic bond-breaking agent tetramethylurea, or incubation in absence of salt, or at low temperature also reduced attachment (P < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)
...
PMID:Adherence of Mycoplasma hyopneumoniae to porcine ciliated respiratory tract cells. 821 93

A Mycoplasma gallisepticum (MG) strain R protein of 64 kilodaltons (p64) was partially digested from the surface of the bacterium by trypsin. Monospecific polyclonal anti-p64 IgG inhibited attachment of MG to chick tracheal rings by as much as 69%. However, trypsin treatment of viable MG cells did not reduce attachment to tracheal rings or hemagglutination titer. Anti-p64 IgG inhibited growth of MG strain R in broth and on solid media, inhibited the uptake of radiolabeled thymidine, but did not inhibit hemagglutination. Anti-p64 IgG inhibited growth of eight MG strains on solid medium. The degree of growth inhibition varied widely depending on the strain and correlated positively with the reported virulence of the MG strains with one exception (A5969). An IgG monoclonal antibody to p64 (MyG 001) inhibited growth of MG strain R on solid and in broth media. The strong attachment-inhibition activity of anti-p64 IgG may result from its growth-inhibiting activity. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of MG strains suggested that p64 is expressed in higher amounts in vitro in virulent strains (R, S6) than in strains of low virulence (F, M876, K503, K703, K730). P64 should be used to immunize chickens to determine if it can stimulate a growth and attachment-inhibiting response in the respiratory tract.
...
PMID:Inhibition of Mycoplasma gallisepticum growth and attachment to chick tracheal rings by antibodies to a 64-kilodalton membrane protein of M. gallisepticum. 825 60

Mycoplasma salivarium cells were demonstrated to bind human IgG Fc fragments. The binding capacity was 78.8% enhanced by incubation of the cell suspension in PBS with 0.25% trypsin at 37 degrees C for 1 h, but tended to fall after incubation with higher concentrations of the enzyme, and was 95.5% lower after incubation of the suspension without trypsin. Fc fragments of IgG from rat, swine, sheep, rabbit, goat, cow and mouse also bound to the organism cells with increasing affinity in this order. The affinity of human IgG Fc fragments was almost comparable with those of sheep and rabbit. Antigen specific IgG from goat (specific for gamma-chain of human IgG, and mu-chain of human IgM) and rabbit (specific for whole molecules of goat IgG) bound to the cells. Binding of goat IgG Fc fragment was inhibited in the presence of antigen specific goat IgG (specific for gamma-chain of human IgG). These results suggest that M. salivarium cells bind IgG from a variety of animal species via the Fc fragment.
...
PMID:Binding of Fc fragments of IgG from human and seven animal species to Mycoplasma salivarium cells. 829 54

The wall-less procaryote Mycoplasma fermentans is currently being examined as an agent potentially associated with human disease, including infectious processes affecting immunocompromised individuals. To delineate and understand the interactions of M. fermentans with its host, specific membrane surface components were characterized as markers for detecting the organism and for assessing heterogeneity in antigenic surface architecture within this mycoplasma species. Detergent phase fractionation of metabolically labeled organisms of type strain PG18 identified a family of prominent integral membrane proteins; several of these labeled with 35S-cysteine and 3H-palmitate, which are characteristics of procaryotic lipoproteins. Specific monoclonal and polyclonal antibodies raised to strain PG18 components further distinguished seven of these membrane proteins, which were localized on the organism's surface by monitoring their selective susceptibility during trypsin treatment of intact cells. With these antibodies, Western immunoblot profiles of surface membrane antigens expressed on strain PG18 were compared with those expressed on the recently identified Incognitus strain of M. fermentans, as well as with several other human and animal mycoplasma species. While the antibodies were specific for M. fermentans, marked differences were observed between the strains in the size of one surface lipoprotein and in the apparent levels of several antigens expressed in the cultured populations analyzed. Some monoclonal antibodies to strain PG18 and a previously described monoclonal antibody to strain Incognitus showed apparent selectivity for the strain used for immunization. Monoclonal antibodies developed here recognize stable epitopes defining a family of surface lipoproteins and provide critical tools to determine the basis of surface variation in this mycoplasma species and to assess the location and antigenic phenotypes of organisms in the human host.
...
PMID:A family of strain-variant surface lipoproteins of Mycoplasma fermentans. 833 64


<< Previous 1 2 3 4 5 6 7 8 9 10 Next >>

---