Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.4.21.4 (trypsin)
 42,187 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

A new non-strain-specific ascites subline of the TA3 mammary adenocarcinoma TA3-MM, which arose in vivo from the strain-specific TA3-St subline during an acute respiratory illness of the syngeneic mouse strain A/HeHa hosts, possessed at its surface a glycoprotein not found on the parent TA3-St cell. This glycoprotein, termed TA3-MM epiglycanin, was characterized by a high molecular weight (500,000), by potent inhibition of hemagglutination by the Vicia gramines lectin, and by carbohydrate and amino acid compositions nearly identical to those of the glycoprotein epiglycanin present at the surface of the allotransplantable TA3-Ha ascites cell. By electron microscopic examination, TA3-MM epiglycanin appeared as long extended rods with widths (2.5 nm) and lengths (450--500 nm) similar to those of TA3-Ha epiglycanin. Incubation of each of two sublines of the TA3-MM ascites cell, TA3-MM/1 and TA3-MM/2, with a modified trypsin followed by column chromatography produced approximately 1.0- and 0.2-fold as much epiglycanin-like material, respectively, as was obtained from the TA3--a ascites cell. Continuous growth of the TA3-MM cell in suspension culture resulted in an almost complete disappearance of epiglycanin in a manner demonstrated earlier for the TA3-Ha cell grown under similar conditions. Allotransplantability in the TA3-MM cell may be due, at least in part, to masking a histocompatibility antigens by epiglycanin-like molecules.
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PMID:Isolation and partial characterization of an epiglycanin-like glycoprotein from a new non-strain-specific subline of TA3 murine mammary adenocarcinoma. 28 25

The preparation of greater than 30 different hybridomas, all secreting IgM class antibodies against epiglycanin, a glycoprotein at the surface of the mouse mammary carcinoma cell line TA3-Ha, is described. The specificities of 10 of the antibodies, with affinity constants in the range of 10(8)-10(10) l/mol were compared in an enzyme competitive binding assay. The affinity of epiglycanin was strongly reduced for all antibodies tested by incubation with periodate (10 mM, 4 degrees C) and was reduced for most of the antibodies by endo-alpha-N-acetyl- D-galactosaminidase. This suggested that carbohydrate, and specifically the Gal beta (1----3)GalNAc disaccharide, formed an integral part of the epitopes of most of the antibodies. The isolated disaccharide, however, exhibited 250,000 times less inhibitory activity in the competitive binding assay than epiglycanin. The binding capacity of epiglycanin was also reduced by incubation with trypsin or pronase, suggesting a high molecular weight dependency for binding. Incubation with sialidase increased its affinity for the antibodies. The binding of the antibodies to epiglycanin was strongly inhibited by peanut agglutinin, and to a lesser extent by lectins from Triticum vulgaris, Ricinus communis, Pisum sativum and Phaseolus vulgaris. None of the antibodies bound to any of eight different gangliosides immobilized on HPTLC plates. Mono- (Fab) and divalent [F(ab')2] fragments of the antibodies possessed very low affinity for epiglycanin. The results demonstrated that the specificities of the antibodies are related, but distinguishable, and they suggest that this epiglycanin-IgM model may be useful for studies on the general principles of the interaction between IgM antibodies and mucin-type glycoproteins.
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PMID:Development and characterization of monoclonal antibodies against a mucin-type glycoprotein. 149 19

Mannose-labeled epiglycanin was prepared by incubation of TA3-Ha ascites cells with [2-3H]mannose, removal of the epiglycanin by incubation of viable cells with L-1-p-tosylamino-2-phenylethyl chloromethyl ketone-trypsin, and isolation of the large epiglycanin glycopeptides by gel filtration. Purification of epiglycanin glycopeptides was performed by wheat germ agglutinin affinity chromatography. Extensive incubation of epiglycanin with Pronase, followed by passage through a calibrated column of Bio-Gel P-4 (Column P-4), gave three fractions. The fraction of lowest apparent molecular weight, about 5000, upon incubation with a purified extract from F. meningosepticum containing an N-glycosyl hydrolase and an endo-N-acetyl-beta-D-glucosaminidase (T.H. Plummer et. al. (1984) J. Biol. Chem. 259, 10700-10704) and passage through Column P-4 gave a peak of radioactivity at apparent Mr 3000. Incubation of nonlabeled epiglycanin under similar conditions with the same enzyme preparation followed by passage through Column P-4, gave two peaks, based upon total mannose content. One of these, partially deglycosylated epiglycanin, was present in the void volume. Its composition indicated that approximately 80% of the mannose content of epiglycanin had been removed by the enzyme treatment, whereas no change was noted in the proportion of the other carbohydrate components. The effluent volume of the second peak coincided precisely with the peak obtained from the Pronase-cleaved fraction. Its composition and apparent Mr were consistent with those of an N-lactosamine-type chain with four antennae, Man3Gal4GlcNAc5NeuAc2-3.
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PMID:Evidence for the presence of an N-acetyllactosamine-type chain in epiglycanin. 378 44