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)

Primary rhesus monkey kidney (MK) cells have long been the cells of choice for isolation and propagation of the human paramyxoviruses (parainfluenza 1, 2, 3, 4A, 4B, and mumps). However, problems with the supply and cost of MK cells and the presence of endogenous viruses, including herpes B virus and SV-5, necessitated a search for an alternative cell line. Continuous cell cultures of human origin (L132, A-549, HuT-292, HEK, G-293, G-401, A-498, A-704, CAKI-1, RD) and simian origin (LLC-MK2, BSC-1, MA-104, Vero) were evaluated for their capacity to support the growth of the human paramyxoviruses, as followed by cytopathic effect, hemadsorption, hemagglutination, and EIA. NCI-H292 (HuT-292) human lung mucoepidermoid carcinoma cells (ATCC # CRL-1848) proved to be the most sensitive line for cultivating all serotypes and strains of the paramyxoviruses. These cells were also shown to be a suitable substitute for MK in primary isolation of paramyxoviruses from clinical specimens. RPMI-1640 with 1.5 micrograms/ml trypsin was the preferred maintenance medium; alternatively, Eagle's MEM supplemented with 1.5 micrograms/ml trypsin and 0.1% ITS was satisfactory. NCI-H292 cells are a continuous line with excellent growth characteristics, although the genetic polyploidy of the cells may limit the number of passages of usable cells.
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PMID:NCI-H292 as an alternative cell line for the isolation and propagation of the human paramyxoviruses. 226 Sep 24

Rotaviruses were isolated following cell culture of the intestinal contents of four non-diarrheic calves. The four isolates were serially propagated in MDBK and BSC-1 cells in the presence of trypsin and produced rotavirus particles morphologically similar to those found associated with diarrhea. They were antigenically related to the Nebraska calf rotavirus (Norden strain) as investigated by immunofluorescence. Three isolates could be distinguished from the reference Nebraska rotavirus by their thermal stability and/or their differential responses to intestinal neutralizing antibodies. Two isolates produced on BSC-1 cells plaques significantly different in size from the reference strain, No significant genomic variations were detected among the isolates.
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PMID:Distinct rotaviruses isolated from asymptomatic calves. 298 33

The ability of mammalian rotaviruses to replicate in BSC-1 and CV-1 cell cultures was facilitated by the presence of 4% chicken serum. Viral plaques tended to be larger and appear more quickly than in cultures without added chicken serum. It is proposed that chicken serum facilitates plaquing of rotavirus due to its lack of trypsin inhibitors.
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PMID:Propagation of rotaviruses in the presence of chicken serum. 608 11

Rotaviruses were isolated on BSC-1 cells from counterimmunoelectrophoresis and/or electron microscopy positive intestinal contents from two asymptomatic and six diarrheic calves from Quebec. The plaque assay was performed using these lines and agar overlay medium containing trypsin and DEAE-dextran. This assay was used to compare the Quebec isolates to an attenuated American strain (NCDV) and another strain (TH) obtained from France. The NCDV strain produced plaques that were significantly larger than those produced by the TH strain. Three Quebec isolates produced plaques similar in size to TH strain, one isolate was similar to NCDV strain and another isolate produced larger plaques than those of both NCDV and TH strains. The other isolates induced the production of plaques that were not significantly different from those of NCDV or TH strains.
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PMID:Comparative study of bovine rotavirus isolates by plaque assay. 608 82

Bovine rotavirus infectivity for continuous green monkey kidney (BSC-1) cells was enhanced in hypertonic medium and following incorporation of cortisol, retinoic acid and vitamin B12 into the cell culture maintenance medium. The virus yields produced under these conditions were similar whether obtained in the presence or absence of trypsin. Infectivity titres were increased following the incorporation of trypsin in the maintenance medium throughout the infection cycle but remained unchanged after trypsin treatment of infected cell extracts. Bovine rotavirus infectivity was not affected by incorporation of phytohaemagglutinin, thyroid gland extracts or foetal calf serum in similar experiments. Unexpectedly, serum promoted virus growth when used with cells treated with actinomycin D. Marked differences in the stability of the newly produced infectious rotavirus were observed, suggesting that cell permissiveness to rotavirus infection may vary following the physiological state of the host cell.
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PMID:Enhanced production of infectious rotavirus in BSC-1 cell cultures by various factors in the presence of absence of trypsin. 626 38

LLC-MK2, GMK AH-1, BSC-1, and Vero cells were compared in titrations of recent isolates and laboratory strains of influenza A and B and parainfluenza types 1, 2, and 3 viruses. About the same titres, as determined by haemadsorption in cell cultures, were obtained in LLC-MK2, GMK AH-1, and BSC-1 cells when trypsin had been added to the medium, whereas the Vero cells were less sensitive to the influenza virus strains tested. Virus titres were usually low in the absence of trypsin. A laboratory strain of parainfluenza 2 virus reached about the same titres in medium without as in medium with trypsin, possibly owing to prior adaptation by passages in Vero cells. Comparative titrations of influenza A, and parainfluenza 1 and 3 viruses suggested the same susceptibility of LLC-MK2 cells with trypsin as of primary monkey kidney cells. Re-isolation experiments from 38 clinical specimens showed LLC-MK2 cells to be as efficient as primary monkey kidney cells for isolation of influenza and parainfluenza viruses, whereas the susceptibility of the other cell lines to clinical material has not yet been tested on a larger scale. It is concluded that a continuous line of monkey kidney cell culture may be acceptable as an alternative to primary monkey kidney cells for the isolation of influenza and parainfluenza viruses from patients.
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PMID:Susceptibility of continuous lines of monkey kidney cells to influenza and parainfluenza viruses in the presence of trypsin. 627 42

Four proteins, GP1, VGP48, GP26 and VPM27, are associated with the envelope of respiratory syncytial (RS) virus. The status of GP1 has been uncertain, because a cellular glycoprotein migrates at the same position when Laemmli's discontinuous buffer system is used for PAGE, and because BSC-1 cells infected with the RSN-2 strain of RS virus appear not to contain GP1. However, additional evidence suggests that GP1 is a viral structural protein. (i) It is removed from cells by trypsin, while the cellular glycoprotein is not; (ii) it is separated from the cellular glycoprotein when the infected cells are analysed by neutral SDS-PAGE; (iii) it is present in the purified RSN-2 strain of RS virus produced by BSC-1 cells; (iv) it is also present in the purified Long strain of RS virus produced by either human or monkey cells. When purified Long strain virus is analysed by PAGE under non-reducing conditions, the glycoproteins VGP48 and GP26 migrate together, and VPM27 separates into two proteins, which one-dimensional peptide mapping suggests are not different proteins. These observations suggest that VGP48 and GP26 exist in the virion as a single molecule joined by disulphide bonds, and so resemble a paramyxovirus fusion protein, and that probably there are two forms of VPM27 which differ in either position or number of disulphide bonds.
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PMID:Respiratory syncytial virus polypeptides. III. The envelope-associated proteins. 683 7

A recent report described a thrombin inhibitory activity in the soluble fraction of human placenta and the cytosolic fraction of K562 cells. Isolation and characterization of the functionally inactive 35-38-kDa placental form of this protein revealed that it was a novel serine proteinase inhibitor (Coughlin, P. B., Tetaz, T., and Salem, H. H. (1993) J. Biol. Chem. 268, 9541-9547). In the present study, we observed a 67-kDa sodium dodecyl sulfate (SDS)-stable complex when 125I-thrombin was incubated with the cytosolic fraction of a monkey kidney epithelial cell line, BSC-1. This complex was not observed in either the particulate cell fraction extracted with 0.2% Triton X-100 or medium conditioned by cells, suggesting that the thrombin-complexing factor is confined to the cytoplasm. The cytoplasmic antithrombin activity was purified to apparent homogeneity from the cytosol of BSC-1 cells previously pulsed with [35S]methionine by a combination of heparin-agarose chromatography, Mono Q fast protein liquid chromatography, and anhydrotrypsin-Affi-Gel 10 affinity chromatography. Analysis of the affinity-purified preparation by SDS-polyacrylamide gel electrophoresis and fluorography revealed a single protein with an apparent molecular mass of 38 kDa. The purified 38-kDa protein inhibited the amidolytic activities of thrombin, trypsin, urokinase, and factor Xa but not that of elastase. Incubation of the 38-kDa protein with excess thrombin identified approximately 60% of the labeled 38-kDa protein in an SDS-stable 67-kDa complex. The purified 38-kDa inhibitor was cleaved with cyanogen bromide and the isolated peptides subjected to microsequencing. Amino acid sequence obtained for a region within this protein exhibited significant homology with human antithrombin III and plasminogen activator inhibitors 1 and 2. This homologous peptide contained the full complement of residues designated as highly conserved in helix F of the greater serine proteinase inhibitor superfamily. In addition, an internal sequence of GGGGDIHQGF was found in the monkey cytoplasmic inhibitor, which is identical to that reported for an internal sequence of the human placental inhibitor. These findings confirm the existence of a novel cytoplasmic serine proteinase inhibitor in mammalian cells and provide additional details of its molecular properties. The physiological function of this novel serine proteinase inhibitor in cytoplasm is unknown.
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PMID:Isolation and characterization of an intracellular serine proteinase inhibitor from a monkey kidney epithelial cell line. 840 7