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)

Transport activity of the hog gastric (H+ + K+)-ATPase system was measured either as the formation of proton gradient using the dye probe acridine orange or as the formation of a proton diffusion potential using the cyanine dye 3,3'-diethyloxdicarbocyanine iodide in the presence of the protonophore tetrachlorosalicylanilide. The development of these gradients has been compared in K+ media in the presence of either Cl- or SO4-2 as the anionic species. This comparison of proton diffusion potential formation to proton gradient formation has been used to demonstrate that a Cl- conductance in this vesicular system results from limited enzymic digestion with either trypsin or alpha-chymotrypsin from the ageing process itself. The possible significance of this finding is discussed.
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PMID:Induction of a chloride conductance in gastric vesicles by limited trypsin or chymotrypsin digestion or ageing. 3 2

A 36-yr-old black female presented with primary amenorrhea. The chromosomal constitution based on QFQ (Q bands by fluorescence using quinacrine) RFA (R bands by fluorescence using acridine orange), GTG (G band by Giemsa using trypsin), and CBG (C band by Giemsa using barium hydroxide) techniques was 46, XX, duplicated (9; q12), inverted (9; p12q12.1) in lymphocytes and skin fibroblasts. Both sex chromosomes were normal. Buccal smear revealed 22% Barr bodies. Duplication and inversion of secondary constriction regions of chromosome 9 may possibly be associated with abnormal clinical features.
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PMID:Primary amenorrhea in a black female with duplication and inversion of the secondary constriction regions of chromosome 9. 12 3

A virus was isolated from the spleen of a white-tailed deer (Odocoileus virginianus) that had died during an epizootic in Washington state in 1967. Inoculation of a 10% spleen suspension from the deer caused hemorrhagic disease in normal white-tailed deer. Studies were conducted on the biological, physicochemical, and serologic properties of the Washington isolate. An in vitro assay system, utilizing a cultured primary of white-tailed deer fetal cells from an entire fetus, was employed for isolation and propagation of the virus. Cytopathic effect was characterized by focal development of rounded and clumped cells. Propagation was unsuccessful in suckling mice, BHK-21, and Vero cell cultures. The virus was resistant to treatment with ether, sodium deoxycholate, trypsin, oxytetracycline hydrochloride, and was sensitive to chloroform. Virus yield was not affected when infected cultures were treated with 5-iodo-2'-deoxyuridine, but dactinomycin (actinomycin D) treatment of infected cultures reduced virus yield. The virus was inactivated when heated at 70 C for 5 minutes or when exposed to pH 5 for 18 hours at 4 C. The virus was completely excluded from the filtrate by a 0.10- micronm (APD) membrane filter. Staining of infected cells with acridine orange indicated the presence of double-standard nucleic acid in the cytoplasm. Serum-neutralization tests with antiserums against the homologous virus and the New Jersey and Alberta strains of epizootic hemorrhagic disease virus resulted in neutralization of the Washington isolate. The Washington virus was not neutralized by bluetongue virus antiserum. Cells infected with the Washington isolate exhibited intracytoplasmic fluorescence by the indirect fluorescent antibody method with New Jersey and Alberta epizootic hemorrhagic disease antiserums but not with bluetongue antiserum.
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PMID:Isolation and characterization of epizootic hemorrhagic disease virus from white-tailed deer (Odocoileus virginianus) in eastern Washington. 19 10

Strains of Streptococcus mutans synthesized bacteriocins in agar plates, but synthesis of detectable bacteriocins in liquid media took place only under certain culture conditions. The composition of the medium proved to be crucial. Trypticase Soy Broth with 4% Yeast Extract meeting the requirements. The effect of the Yeast Extract is obscure, for some strains also formed detectable bacteriocins in a special Trypticase medium without this agent. It was noted that the broth should be filter-sterilized rather than autoclaved and only a few days old. Attempts at liberating cell-bound bacteriocins from washed cells were unsuccessful, even when they were treated with ultrasound, EDTA, or various chemicals followed by ultrasound. On the basis of size and sensitivity to heat the bacteriocins could be divided into two groups, while their resistance to ether and chloroform and to trypsin did not follow this pattern. Dependence on plasmids could not be demonstrated by attempts at curing with acridine orange or ethidium bromide; and the involvement of phages was unlikely, since the inhibition was not transmissible and phage-like structures were not observed in the electron microscope.
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PMID:Synthesis of bacteriocins in liquid cultures of Streptococcus mutans. 26 10

A marked increase in water permeability can be induced in Xenopus oocytes by injection of mRNA from tissues that express water channels, suggesting that the water channel is a protein. In view of this and previous reports which showed that proteinases may interfere with mercurial inhibition of water transport in red blood cells (RBC), we examined the influence of trypsin, chymotrypsin, papain, pronase, subtilisin and thermolysin on water permeability as well as on ATPase activity, H(+)-pump, passive H+ conductance, and Na+/H+ exchange in apical brush-border vesicles (BBMV) and endosomal (EV) vesicles from rat renal cortex. H+ transport was measured by Acridine orange fluorescence quenching and water transport by stopped-flow light scattering. As measured by potential-driven H+ accumulation in BBMV and EV, proteinase treatment had little effect on vesicle integrity. In BBMV, ecto-ATPase activity was inhibited by 15-30%, Na+/H+ exchange by 20-55%, and H+ conductance was unchanged. Osmotic water permeability (Pf) was 570 microns/s and was inhibited 85-90% by 0.6 mM HgCl2; proteinase treatment did not affect Pf or the HgCl2 inhibition. In EV, NEM-sensitive H+ accumulation and ATPase activity were inhibited by greater than 95%. Pf (140 microns/s) and HgCl2 inhibition (75-85%) were not influenced by proteinase treatment. SDS-PAGE showed selective digestion of multiple polypeptides by proteinases. These results confirm the presence of water channels in BBMV and EV and demonstrate selective inhibition of ATPase function and Na+/H+ exchange by proteinase digestion. The lack of effect of proteinases on water transport by mercurials. We conclude that the water channel may be a small integral membrane protein which, unlike the H(+)-ATPase and Na+/H+ exchanger, has no functionally important membrane domains that are sensitive to proteolysis.
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PMID:Proteinases inhibit H(+)-ATPase and Na+/H+ exchange but not water transport in apical and endosomal membranes from rat proximal tubule. 130 58

Intrahepatic bile duct epithelial cells, or cholangiocytes, contribute to bile secretion in response to hormones, including secretin. However, the mechanism by which secretin stimulates ductular bile flow is unknown. Since recent data in nonhepatic epithelia have suggested a role for exocytosis in fluid secretion, we tested the hypothesis that secretin stimulates exocytosis by isolated cholangiocytes. Cholangiocytes were isolated from normal rat liver by a newly described method employing enzymatic digestion and mechanical disruption followed by immunomagnetic separation using specific monoclonal antibodies, and exocytosis was measured using a fluorescence unquenching assay employing acridine orange. Secretin caused a dose-dependent (10(-12)-10(-7) M) increase in acridine orange fluorescence by acridine orange-loaded cholangiocytes with a peak response at 10 min; the half-maximal concentration of secretin was 7 x 10(-9) M. The secretin effect was inhibited by preincubation of cholangiocytes with colchicine (30% inhibition, p less than 0.05) or trypsin (90% inhibition, p less than 0.001); no inhibition was seen with lumicolchicine and heat-inactivated trypsin. Cholecystokinin, insulin, and somatostatin had no effect on fluorescence of acridine orange-loaded cholangiocytes; secretin had no effect on fluorescence of acridine orange-loaded hepatocytes or hepatic endothelial cells. Exposure of isolated cholangiocytes to secretin at doses that stimulated exocytosis caused a dose-dependent increase in cyclic AMP levels (218% maximal increase, p less than 0.05); moreover, an analogue of cyclic AMP stimulated exocytosis by cholangiocytes. Secretin had no effect on intracellular calcium concentration using Fura-2-loaded cholangiocytes assessed by digitized video microscopy. Our results demonstrate, for the first time, that secretin stimulates exocytosis by rat cholangiocytes. The effect is cell- and hormone-specific, dependent on intact microtubules, on a protein(s) on the external surface of cholangiocytes, and on changes in cellular levels of cyclic AMP. The results are consistent with the hypothesis that secretin-induced changes in bile flow may involve an exocytic process.
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PMID:Secretin stimulates exocytosis in isolated bile duct epithelial cells by a cyclic AMP-mediated mechanism. 132

Early and late replicating chromosomal banding patterns of Gallus domesticus were investigated by cell synchronization and incorporation of 5'-bromodeoxyuridine during early and late DNA synthesis. The early replicating chromosomal banding patterns observed, as revealed by either acridine orange or Hoechst 33258/propidium iodide staining, were similar to the structural G-banding patterns obtained by trypsin digestion and Giemsa staining. Late replicating chromosomal banding showed extensive reverse band complementarity to the G-banding pattern. Cell synchronization increased the number of prometaphase and metaphase plates available for analysis. G-banding obtained by Hoechst 33258/propidium iodide staining was investigated due to the fact that it is compatible with chromosomal in situ hybridization procedures that use nonisotopically-labeled DNA probes. Standard replicative G-banded and R-banded idiograms, as obtained after cell synchronization, are proposed.
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PMID:Early and late replicative chromosomal banding patterns of Gallus domesticus. 137 28

An acridine derivative of N-alpha-Fmoc-lysine has been prepared and used in solid phase peptide synthesis. The fluorescence properties of the acridine reporter group are retained throughout the peptide synthesis procedure. The utility of the acridine group was demonstrated by its use as an energy acceptor in a fluorescence energy-quenching assay with trypsin.
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PMID:An acridine amino acid derivative for use in Fmoc peptide synthesis. 158 39

Hydrocortisone affected less distinctly the gene expression in rat liver slices as compared with the simultaneous effect of the hormone and blood plasma high density lipoproteins (HDL). These substances acted cooperatively and the phenomenon observed was not reproduced by means of individual components. The protein moiety of HDL, but not of lipid part of the molecule, was of importance in realization of the effect. Inhibitors of lysosomal proteinases (pepstatin, soy bean inhibitor of trypsin and iodoacetamide) as well as inhibitors of lysosomes intracellular translocation (colchicine and vinblastine) blocked completely the cooperative action of hydrocortisone and HDL. Hydrocortisone-dependent activation of chromatin and stimulation of genes expression led to an increase in the binding constant of acridine orange with DNA, while molar concentration of binding sites of the probe was practically unaltered. At the same time, simultaneous effect of hydrocortisone and HDL did not alter the constants for acridine orange binding but concentration of the probe binding sites with DNA was distinctly increased.
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PMID:[The role of plasma high density lipoproteins as modulators of specific hormonal effect of hydrocortisone]. 238 32

Streptococcus faecalis has been reported to cause food poisoning. Six strains of S. faecalis were tested for Sherman's criteria. These strains were non-hemolytic, DNase+ and Ent+. The enterotoxin was purified on Sephadex G-200 column and maximum activity was observed at 37 C and pH 7.0. Enterotoxin treated with trypsin and papain elicited very poor response to fluid accumulation. The sensitivity of all the strains against different antibiotics was determined. Strain 53 M was treated with acridine orange and ethidium bromide and a total of 44 Amps Strr and 3 Amps Strs mutants were tested for toxin production. Out of these only 4 were toxin negative, amongst which 3 were also DNase negative and 1 showed partial DNase activity.
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PMID:Studies on Streptococcus faecalis enterotoxin. 244 76


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