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

An adherent cell line, termed TC-1, has been isolated from long-term liquid culture of murine marrow cells by repeated exposure of the adherent cells to 0.1% trypsin. This is an alkaline phosphatase-positive cell line showing variable staining with acid phosphatase and alpha-naphthyl acetate esterase. On electron microscopy, the cells have moderate amounts of rough endoplasmic reticulum and variable numbers of polyribosomes. Some cells contain large clusters of laked glycogen particles. Intermediate junctions are present between some cells. Conditioned medium from this cell line produced from 384 to 638 units of CSF-1 per milliliter by radioimmunoassay and a CSF-1-dependent synergistic activity, which stimulates giant macrophage colony formation of marrow cells in soft agar. The conditioned medium also stimulates 3H-TdR incorporation by marrow cells in liquid culture and induces secondary adherent cell lines. The growth factor(s) produced by the TC-1 stromal cell line may be important in the regulation of early stages of hematopoietic differentiation. Two subclones, TC-1-C-11 and TC-1-C-3, have been isolated from passage 25 of the TC-1 cells by a penicylinder separation technique. The TC-1-C-11 is phenotypically like the parent TC-1 line and produces macrophage growth factors. The TC-1-C-3 grows as an epithelioid monolayer with visible junctions among adjacent cells under phase contrast microscopy. This subclone produces retrovirus and is capable of providing anchorage support for hematopoietic stem cells. The TC-1 cell line and its subclones may provide models for the control of early stem cell proliferation and differentiation.
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PMID:Hematopoietic factor production by a cell line (TC-1) derived from adherent murine marrow cells. 241 62

Circulating concentrations of digestive enzymes, certain lysosomal hydrolases and protease inhibitors were measured in 19 heavy smokers and 13 non-smokers before (basal) and at 15, 30, and 60 minutes after a single intravenous injection of secretin (75 CU). In smokers, basal serum amylase and immunoreactive pancreatic elastase 2 (IRE2) concentrations were about 100% and 25% higher respectively, than in the non-smokers, whereas, no differences were observed in basal immunoreactive cationic trypsinogen (IRCT) concentrations and in acid phosphatase and beta-glucuronidase activities between the two groups. Furthermore, a single injection of secretin to cigarette smokers significantly increased serum amylase, IRCT and IRE2 by 155%, 200%, and 100%, respectively when compared with their corresponding basal levels. No such increment was observed in the non-smokers. In addition, there were no significant differences in serum trypsin or elastase inhibitory capacity or immunoreactive alpha 1-protease inhibitor and alpha 2-macroglobulin levels between smokers and non-smokers. The levels and inhibitory capacity of these protease inhibitors was also not affected by secretin injection. These data suggest that cigarette smoking enhances the responsiveness of the exocrine pancreas to a physiological stimulus such as secretin, with resultant substantial increase in the concentrations of pancreatic hydrolases in blood.
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PMID:Raised serum concentrations of pancreatic enzymes in cigarette smokers. 243 81

In an effort to develop a model of chronic alcoholic pancreatitis in Sprague-Dawley rats fed a nutritionally adequate diet, 3 groups of 15 animals each were fed Wayne Rodent-Blox ad libitum, Lieber-DeCarli diet with 40% of carbohydrate calories replaced by ethanol ad libitum and isocaloric amounts of Lieber-DeCarli diet respectively for a period of 18 months. Rats were anesthetized and basal and secretin-stimulated pancreatic juice was obtained. Pancreatic glands were isolated and divided into portions for histology, biochemical analyses, and cell fractionation. The homogenate, zymogen granule fraction, mitochondrial-lysosomal fraction, microsomal fraction and postmicrosomal supernatant as well as aliquots of pancreatic juice were analyzed for cathepsin B, acid phosphatase, beta-D-glucoronidase, arylsulphatase and leucine naphthylamidase. All of the ethanol-fed animals developed morphological changes akin to human chronic pancreatitis. There were focal areas of parenchymal degeneration with fibrosis, protein plug formation and tubular complexes. In the pancreatic tissue of animals fed ethanol, total protein, trypsinogen (and free trypsin) were increased and amylase was decreased. While acid phosphatase was increased in all of the particulate fractions, cathepsin B was increased in the zymogen granule and mitochondrial-lysosomal fractions. Basal and post-secretin pancreatic juice did not show a significant increase in digestive or lysosomal enzymes. It is suggested that focal degenerative changes may be due to trypsin generated by intracellular activation of digestive enzymes by lysosomal enzyme cathepsin B.
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PMID:Alcoholic pancreatitis in rats fed ethanol in a nutritionally adequate liquid diet. 244 6

Human prostatic acid phosphatase is known to display considerable charge heterogeneity upon isoelectric focusing. The structural basis of this heterogeneity is not known, although it has been widely attributed to variations in the nature of the carbohydrate chains or to substituents on the carbohydrate chains of the glycoprotein. In this study, the role of the carbohydrate chains in the charge heterogeneity of the protein was examined. First, sialic acid residues were removed by treatment of the acid phosphatase with neuraminidase. The desialo enzyme was fractionated and purified by L-tartramic acid affinity chromatography. Then, after the protein oligosaccharide linkages were made accessible by the presence of NP-40 or by denaturing the protein, the protein was completely deglycosylated by endo-beta-N-acetylglucosaminidase F at pH 4.5 and 9.3. Two discrete intermediates were clearly resolved by SDS gel electrophoresis during the deglycosylation of the denatured protein at pH 9.3, indicating the existence of three sites of glycosylation on the protein. Peptide mixtures were obtained by digestion of carboxymethylated and citraconylated derivatives of the enzyme with trypsin and the glycopeptides were isolated. The amino acid compositions of the glycopeptides were consistent with the interpretation that there are a minimum of two sites of glycosylation on each peptide subunit of the enzyme. Isoelectric focusing experiments on the native, desialo, and denatured, deglycoso acid phosphatase showed that the heterogeneity of the protein is not eliminated either by desialylation or by deglycosylation. Thus, the electrophoretic heterogeneity of human prostatic acid phosphatase does not lie primarily in the oligosaccharide part of the glycoprotein or in altered conformational states of the protein, but in structural variations of the polypeptide itself. The heterogeneity may be due to variations at the C-terminus, partial deamidation, phosphorylation, sulfation or other posttranslational modifications of the protein chain.
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PMID:Carbohydrate removal fails to eliminate the heterogeneity of human prostatic acid phosphatase. 250 33

The endoplasmic reticulum-localized enzyme alpha-glucosidase II is responsible for removing the two alpha-1,3-linked glucose residues from N-linked oligosaccharides of glycoproteins. This activity is missing in the modA mutant strain, M31, of Dictyostelium discoideum. Results from both radiolabeled pulse-chase and subcellular fractionation experiments indicate that this deficiency did not prevent intracellular transport and proteolytic processing of the lysosomal enzymes, alpha-mannosidase and beta-glucosidase. However, the rate at which the glucosylated precursors left the rough endoplasmic reticulum was several-fold slower than the rate at which the wild-type precursors left this compartment. Retention of glucose residues did not disrupt the binding of the precursor forms of the enzymes with intracellular membranes, indicating that the delay in movement of proteins from the ER did not result from lack of association with membranes. However, the mutant alpha-mannosidase precursor contained more trypsin-sensitive sites than did the wild-type precursor, suggesting that improper folding of precursor molecules might account for the slow rate of transport to the Golgi complex. Percoll density gradient fractionation of extracts prepared from M31 cells indicated that the proteolytically processed mature forms of alpha-mannosidase and beta-glucosidase were localized to lysosomes. Finally, the mutation in M31 may have other, more dramatic, effects on the lysosomal system since two enzymes, N-acetylglucosaminidase and acid phosphatase, were secreted much less efficiently from lysosomal compartments by the mutant strain.
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PMID:Biogenesis of lysosomal enzymes in the alpha-glucosidase II-deficient modA mutant of Dictyostelium discoideum: retention of alpha-1,3-linked glucose on N-linked oligosaccharides delays intracellular transport but does not alter sorting of alpha-mannosidase or beta-glucosidase. 250 71

Since they are found to be increased in lesions of acute necrotic ulcerative gingivitis or marginal periodontitis, agents for these diseases. In the present study, 38 pure cultured strains were obtained as a result of isolation and culture of samples collected from lesions of marginal periodontitis (periodontal pokets), and the biological and biochemical characteristics of these strains were investigated. 1) Light microscopy (including dark-field microscopy) and transmission electron microscopy (negative staining) were used for observation of the morphology and cellular structure of the strains. The cells had a spiral shape, and showed active movement. Based on the above findings the cultured strains were all confirmed to be spirochetes of small to medium size, being 0.08-0.24 micron in width. 2) Growth and motility of the strains were investigated on various types of culture medium. Intense growth and movement were noted in strains cultured in bovine liver exudate medium containing horse serum (pH 7.2) at 37 degrees C under anaerobic conditions produced by the evacuation-replacement method (95% N2, 5% CO2) for 3-7 days after inoculation. 3) Thirty-five strains were positive for indole production and decomposition of urea, mucin, hippuric acid and esculin. Production of hydrogen sulfied was observed in 31 strains. In decomposition tests for 17 carbohydrates, 17 strains were positive for galactose and 14 strains were positive for glucose, while 11 strains were positive for dextrin and 10 strains for fructose upon decomposition of soluble starch. Other carbohydrates were also decomposed by a few strains. 4) In an investigation of the production of alcohol and lower fatty acids, among the metabolic products detected by gas chromatography, a large amount of acetic acid and small amounts of ethanol, lactic acid, propionic acid, pyruvic acid were observed. 5) The results of enzyme activity tests using an API ZYM system indicated relatively high activities of esterase, esterase-lipase, alpha-glucosidase, alkaline phosphatase, trypsin and acid phosphatase.
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PMID:[Biological and biochemical characteristics of the oral spirochetes isolated from the focus of marginal periodontitis]. 276 48

2-(4-Bromo-2,3-dioxobutylthio)-1,N(6)-ethenoadenosine 2',5'-bisphosphate (2-BDB-T epsilon A-2',5'-DP) is an affinity label for the coenzyme-binding site of pig heart NADP+-dependent isocitrate dehydrogenase. Specific reaction occurs at the coenzyme site with an incorporation of 0.5 mol of reagent/mol of enzyme subunit (i.e. modification of only one subunit of the dimeric enzyme) (Bailey, J.M., and Colman, R.F. (1985) Biochemistry 24, 5367-5377). Modified enzyme, prepared by incubating 1 mg/ml isocitrate dehydrogenase with 75 microM 2-BDB-T epsilon A-2',5'-DP in the absence and presence of substrate or coenzyme, was reduced with NaBH4, carboxymethylated, and digested with trypsin. Nucleotidyl peptides were isolated by chromatography on DEAE-cellulose, followed by treatment with acid phosphatase (to decrease the negative charge by removing the phosphate groups from covalently bound reagent) and rechromatography on the same DEAE-cellulose column. The isolated peptides were characterized by amino acid analysis, dansylation, and gas-phase sequencing. A single triskaidekapeptide corresponding to modification of the coenzyme site by 2-BDB-T epsilon A-2',5'-DP was identified as: Asp-Leu-Ala-Gly-X-Ile-His-Gly-Leu-Ser-Asn-Val-Lys. Additional evidence indicated that X is a glutamate residue derivatized by 2-BDB-T epsilon A-2',5'-DP.
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PMID:Isolation of the glutamyl peptide labeled by the nucleotide analogue 2-(4-bromo-2,3-dioxobutylthio)-1,N(6)-ethenoadenosine 2',5'-biphosphate in the active site of NADP+-specific isocitrate dehydrogenase. 288 70

The biosynthesis of distinct prostatic and lysosomal acid phosphatases is demonstrated using a human prostatic carcinoma cell line, PC-3SF12. The biosynthesis and maturation of the acid phosphatases was studied by metabolic labeling with radioactive leucine, specific immunoprecipitation, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and fluorography. Of the tartrate-inhibitable acid phosphatase activity in PC-3SF12 cells, 60% is lysosomal and 10% is prostatic. The lysosomal-type acid phosphatase is synthesized as precursor with a molecular weight of 68,000, some of which is converted to higher-molecular-weight precursor polypeptides (Mr 71,000 and 77,000). The multiple forms of the precursors are due to differences in the carbohydrate chains on the enzyme because biosynthesis in the presence of tunicamycin eliminates the precursor multiplicity. The initial precursor (Mr 68,000) is processed to a mature polypeptide (Mr 49,000), via intermediates with molecular weights of 62,000 and 59,000. The mature polypeptide is degraded to smaller polypeptides with molecular weights of 30,000, 28,000, and 25,000. Precursor polypeptides of the lysosomal-type enzyme are secreted in the medium. Prostatic acid phosphatase is synthesized as a precursor with a molecular weight of 110,000, which is processed via several intermediates (Mr 99,000-93,000, 77,000, and 55,000) to a mature polypeptide with a molecular weight of 49,000. Particularly during cell homogenization, or lysis, the mature polypeptide is rapidly degraded to an immunoprecipitable polypeptide with a molecular weight of 20,000. None of these polypeptides is secreted in detectable amounts into the medium. Precursors and mature and smaller polypeptides are present in human prostate extract and seminal fluid. Proteolytic degradation of prostatic acid phosphatases in cells and tissues is probably catalyzed by a plasmin-like or related trypsin-like enzyme because degradation of the mature prostatic phosphatase polypeptide is completely prevented by addition of the plasmin inhibitor bovine pancreatic trypsin inhibitor. Prostatic- and lysosomal-type acid phosphatases are eventually stored at least in part in two different types of cell organelles. Testosterone does not influence the biosynthesis and secretion of either acid phosphatase in this cell line.
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PMID:Biosynthesis and processing of prostatic and lysosomal acid phosphatases in a prostate carcinoma cell line PC-3SF12. 294 40

Blast cells from ten patients (seven adults, three children) with acute lymphoblastic leukemias (ALL) contained immunoreactive cytoplasmic alpha-1-anti-trypsin (alpha-1-AT) and alpha-1-antichymotrypsin (alpha-1-ACT). Cytochemically positive reactions for block-like periodic acid-Schiff and a localized acid phosphatase suggested that the cells were of lymphoid origin rather than myeloid origin: negative for sudan black-B, nonspecific esterase, chloroacetate esterase, and myeloperoxidase. By surface phenotype, the leukemia showed positive reactions for both lymphoid (common acute lymphoblastic leukemia antigen, Ia, OKT-10) and myeloid (OKM-1, Leu M-1) antigens. Three of three patients tested portrayed the Philadelphia chromosome. Nine patients were Mexican-American and one was Japanese: all were of Asian ethnic derivation. Both myeloid and lymphoid treatment regimens were employed, with survival less than expected. Early granulocytic differentiation detectable by cytoplasmic alpha-1-AT and alpha-1-ACT in lymphoid blasts is discussed.
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PMID:Acute leukemias with both myeloid and lymphoid surface markers. Cytoplasmic alpha-1-anti-chymotrypsin and alpha-1-anti-trypsin as possible indicators of early granulocytic differentiation. 294 24

Renin granules were isolated from rat kidney cortex by a continuous polyvinyl-pyrrolidone-coated colloidal silica (Percoll) density gradient centrifugation. A major peak of renin activity was found at a density of 1.12-1.13 g/ml, and the specific activity of renin in the peak fraction was increased by approximately 70-fold, as compared with that in the kidney cortex homogenate. On the other hand, activities of other reference enzymes, such as succinate dehydrogenase, acid phosphatase and glucose-6-phosphatase, were not detectable in the peak fraction. When the extract of the peak fraction was applied to a pepstatin column, trypsin-activated renin could not be detected in the breakthrough fractions. These results indicate that renin granules of the rat kidney cortex contain only active renin.
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PMID:Renin granules isolated from rat kidney cortex by continuous colloidal silica (Percoll) density gradient centrifugation. 301 76


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