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

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Query: EC:3.4.24.3 (collagenase)
18,340 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Mycobacterium ulcerans produces an exotoxin in culture which, when inoculated into guinea pig skin, causes inflammation, necrosis, edema, and other histopathological changes resembling those in infections of humans. The toxin was resistant to heat and to alkalies and was moderately acid labile. Toxic activity was destroyed by Pronase, phospholipase, lipase, amylase, and glucosidase but not by trypsin, collagenase, cellulase, lysozyme, hyaluronidase, or neuraminidase. Toxic activity was resistant to treatment with 2-mercaptoethanol, urea, guanidine hydrochloride, p-chloromercuribenzoate, ethylenediaminetetraacetate, and sodium deoxycholate but was destroyed by sodium m-periodate and sodium dodecyl sulfate. The toxin was precipitated by a wide range of ammonium sulfate concentrations. Extraction with chlorofrom-methanol or petroleum ether destroyed its activity. Isopycnic density gradient ultracentrifugation in KBr produced a high-density lipoprotein layer with a 24-fold increase in specific activity. The results indicate that this toxin is a high-molecular-weight phospholipoprotein-polysaccharide complex.
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PMID:Further characterization of Mycobacterium ulcerans toxin. 3 Jun 94

A method of dissociation of the rat heart cell for the mixed lymphocyte culture purpose is described. The treatment of the young rat heart with 0.1% collagenase-hyaluronidase solution yielded satisfactory heart cell suspension. The hear cells, thus obtained after mitomycin C treatment but not irradiation, stimulated allogeneic lymphocytes in the presence of 2-mercaptoethanol. In the Fischer to Lewis combination compatible at the Ag-B locus, strong reactions of Lewis lymphocytes to the dissociated heart and skin cells of the Fischer rat were related to the acute rejection of heart and skin grafts, while negative reactions by the kidney and spleen cells reflected a prolonged survival of the kidney graft. The role of skin- and hear-specific antigens in the rejection phenomenon is discussed.
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PMID:Mixed heart cell-lymphocyte reactions and graft survivals in Ag-B-compatible rats. 13 26

Rat liver cells prepared by collagenase treatment and separated by density gradient centrifugation with Ficoll-Hypaque solution (specific gravity 1.120), containing only hepatocytes (80%) and Kupffer cells (20%), were found to stimulate strongly Ag-B-incompatible lumphocytes without 2-mercaptoethanol in culture. The same liver cells could also stimulate Ag-B-compatible lymphocytes in the presence of 2-mercaptoethanol. A positive response was only seen when the liver cell numbers were 1 approximately 2% of the responding lymphocytes. Viable liver cells inhibited the reaction in a two-way culture.
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PMID:Liver and immune responses. IV. Characteristics of the liver cell-lymphocyte interaction. 14 29

Basement membrane procollagen biosynthesis was studied in organ cultures of embryonic rat parietal yolk sac endoderm by following [14C]proline incorporation into nondialyzable proteins. After reduction with 2-mercaptoethanol the 14C-proteins synthesized were characterized by agarose gel filtration and disc electrophoresis in the presence of sodium dodecyl sulfate. The labeled procollagen was identified by its content of hydroxy[14C]proline, its sensitivity to digestion with bacterial collagenase, and its resistance to digestion with pepsin. In cultures which were continuously labeled for periods from 6 hours to 4 days, the pro-alpha chains consistently eluted as a single peak with an apparent molecular weight of 160,000. After pepsin digestion the resultant alpha chains had an apparent molecular weight between 125,000 and 140,000. This suggests that basement membrane procollagen either contains non-triple helical pepsin-resistant regions or a triple helical region which is larger than the corresponding region of interstitial procollagen. Two experiments were performed to determine whether the chains of newly synthesized basement membrane procollagen were cleaved to a smaller molecular species. In the first, the hydroxylation and secretion of procollagen were blocked with alpha, alpha'-dipyridyl, and the resulting intracellular chains of basement membrane protocollagen were found to co-elute with fully hydroxylated and secreted pro-alpha chains. In the second, cultures were labeled for 1 day and chased for 3 days with unlabeled medium. Autoradiography had shown that most of the label was chased into new basement membrane. Agarose chromotography showed that after 3-day chase the pro-alpha chains still eluted with an apparent molecular weight of 160,000. Thus, the data indicated that basement membrane procollagen was deposited in new basement membrane without undergoing a time-dependent extracellular conversion.
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PMID:Basement membrane procollagen is not converted to collagen in organ cultures of parietal yolk sac endoderm. 17 63

Proteoglycan complexes from collagenase [EC 3.4.24.3]-indigestible materials of bovine heart valves were extracted with 4 M guanidinium chloride, purified by ion-exchange column chromatography in a urea-containing solution, then fractionated by density-gradient centrifugation under dissociative conditions. Electrophoretic characteristics and enzymic susceptibility of the density-gradient fractions revealed that the glycosaminoglycans constituting the proteoglycan complexes in this indigestible materials were mainly dermatan sulfate in the top three fractions, and dermatan sulfate and chondroitin sulfates in the bottom fraction; a minor constituent which was common to all the fractions was hyaluronic acid. A gel-like substance (Fr. Ig) at the top of the gradient, amounting to about 25% of the loaded dry sample, contained only a trace of hydroxyproline (less than 1%) and was composed of proteodermatan sulfate, glycoprotein, and a small amount of hyaluronic acid. Sodium dodecyl sulfate-polyacrylamide gel electrophoretic analyses of Fr. Ig with 2-mercaptoethanol showed that the major part of the proteins in this gel-like substance was cross-linked by disulfide bridges. Chromatography of Fr. Ig on Sepharose 4B in buffered 4 M guanidinium chloride containing 2-mercaptoethanol, together with the electrophoretic patterns of the resulting fractions, suggested that proteodermatan sulfate was not associated with hyaluronic acid through covalent bonds. The amino acid composition of Fr. Ig was very similar to that reported in the literature for "dermatan sulfate-protein complex", and "structural glycoprotein" or "acidic structural protein".
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PMID:Proteoglycan complexes from bovine heart valve. Fractionation by density-gradient centrifugation and gel filtration under dissociative conditions. 18 67

A chromatographic method is developed for quantitative estimation of the collagenase-like enzyme (CLE) activity in extract of adenohypophysis and in preparations obtained during various steps of the enzyme isolation. The enzymatic hydrolysis of Cbz-Gly-Pro-Ala-Gly-Pro-Gly-OCH3 in presence of 2-mercaptoethanol at pH 8.0 was used as a pattern. The products formed were separated by chromatography on the paper; then they were stained with ninhydrin and converted into cupric complexes during extraction with ethanol; the optic density was measured at 510 nm. The optimal conditions for the enzymatic reaction were established. The method enabled to estimate the CLE activity in presence of prolyl carboxypeptidase. The specific effect of the CLE purified preparation on various synthetic peptides is discussed.
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PMID:[Chromatographic method of determining the activity of the collagenase-like enzyme of the adenohypophysis and several findings concerning the specificity of its action]. 88 63

The cuticles from distinct developmental stages of Ascaris suum were isolated by a combination of mechanical disruption and detergent treatment of larvae or by surgical removal of cuticle from adults. Proteins from the isolated cuticles were solubilized with SDS and 2-mercaptoethanol (2ME) and analyzed by PAGE. Cuticular proteins from the third and fourth larval stages (L3 and L4) were comparable to adult, but differences in the number of bands were observed. The soluble proteins from the adult, L3 and L4 were readily degraded by bacterial collagenase, suggesting that these proteins are collagen-like structural elements of the cuticle. The soluble proteins from the L2 differed from the adult and other larval stages in both the number and molecular weight of protein bands and their lack of collagenase sensitivity. Antibodies made against the soluble cuticular proteins reacted with the medial and basal layers of the cuticle but not the external cortical or epicuticular regions. A significant amount of the cuticle was not solubilized by 2ME and was not digested by bacterial collagenase. These insoluble cuticular proteins were probably derived from the epicuticular and external cortical regions of the cuticle. Different developmental stages of A. suum were biotinylated and examined by electron microscopy. An organic soluble biotin reagent labeled all stages in a transcuticular pattern, while an aqueous soluble biotin labeled only the external cortical and epicuticular regions of the L4 and adult cuticle. These data indicate the presence of a hydrophobic barrier in the cuticle of later stages of the parasite.
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PMID:The cuticular biology in developmental stages of Ascaris suum. 197 29

Latent and active 58-kDa forms of human neutrophil collagenase (HNC) have been purified to homogeneity. Buffy coats were extracted in the presence and absence of phenylmethanesulfonyl fluoride to generate crude starting preparations that contained latent and active HNC, respectively. The buffers used in preparing these extracts and for all subsequent chromatographic steps contained NaCl at a concentration of 0.5 M or greater, 0.05% Brij-35, concentrations of CaCl2 of 5 mM or greater, and (when feasible) 50 microM ZnSO4 to stabilize the HNC. The collagenase activity in the buffy coat extracts was adsorbed to a Reactive Red 120-agarose column at pH 7.5 in 0.5 M NaCl and was eluted when the NaCl concentration was increased to 1 M. The active and p-(chloromercuri)benzoate-activated latent enzymes were next adsorbed to a Sepharose-CH-Pro-Leu-Gly-NHOH affinity resin in 1 M NaCl at pH 7.5 and desorbed at pH 9 to give a fraction containing only HNC and a small amount of neutrophil gelatinase. The latter enzyme was removed by passage over a gelatin-Sepharose column in 1 M NaCl at pH 7.5. The purified samples of active and latent HNC were obtained with typical cumulative yields of 32 and 82% and specific activities toward soluble rat type I collagen at 30 degrees C of 7200 and 12,000 micrograms min-1 mg-1, respectively. These specific activities are markedly higher than previously reported for HNC. Both active and latent HNC exhibit a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis both in the presence and in the absence of 2-mercaptoethanol. The mobility of latent HNC is consistent with a molecular weight of approximately 58K, with the active form exhibiting a slightly lower (less than 1-2K) molecular weight.
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PMID:Purification to homogeneity of latent and active 58-kilodalton forms of human neutrophil collagenase. 217 75

Protein species found in soluble crude extracts of Hypoderma lineatum (common cattle grub) 1st-instar larvae (HL1) were separated by non-denaturing and denaturing polyacrylamide gel electrophoresis (PAGE) and analyzed for antigenicity by Western blotting using serum from H. lineatum-infested and vaccinated cattle. All HL1 proteins resolved by non-denaturing PAGE were found to be antigenic in the infested bovine host. Treatment of the proteins with sodium dodecyl sulfate and 2-mercaptoethanol destroyed the ability of hypodermin B and the Peak 2 proteins from DEAE-ion exchange HPLC to be bound by antibody. The principal proteins, hypodermin A and hypodermin C (collagenase), appear to be the most immunogenic of the larval proteins. Although having similar amino acid composition, hypodermin A did not appear to share an antigenic epitope with the most prevalent protein, hypodermin C. These results may allow for the selection of proteins to be used in vaccine trials and studies of protective immunological mechanisms associated with acquired resistance to H. lineatum infestation in the bovine host.
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PMID:Antigenicity and immunogenicity of Hypoderma lineatum soluble proteins in the bovine host. 245 35

Cuticular surface antigens of the XL3 and L4 stages of Haemonchus contortus have been studied by surface labeling and immunological techniques. Live worms were labeled with 125I and extracted with sodium dodecyl sulfate (SDS) followed by SDS + 2-mercaptoethanol. The SDS-soluble surface proteins of XL3s and L4s were found to consist of relatively few major species. The pattern of labeled polypeptides was distinctive for each developmental stage. These proteins are refractory to digestion by bacterial collagenase. Several of the proteins are glycosylated. Further extraction of labeled worms with SDS + 2-mercaptoethanol solubilized additional labeled proteins that appeared to be primarily collagens. Rabbit antisera prepared against native XL3 and L4-cuticles reacted strongly with the surfaces of live worms in immunofluorescence assays. In contrast, antisera prepared against SDS-extracted cuticles reacted weakly or not at all with live worms in similar experiments. Rabbit antisera prepared against adult cuticles failed to react with live XL3s or L4s. These studies suggest that the major surface antigens of XL3s and L4s are solubilized by SDS and that there are different antigens present on the cuticular surfaces of XL3s, L4s and adults. Stage-specificity in cuticular surface proteins may contribute to the successful parasitic lifestyle of this nematode.
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PMID:Identification and preliminary characterization of cuticular surface proteins of Haemonchus contortus. 255 12


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