Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.4.21.9 (enterokinase)
 675 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Jejunal perfusion in the rat with Ringer solution containing 10 mmol/l taurocholate removes considerable quantities of protein and brush border membrane enzymes from the intestinal epithelium. The duration of the experiments was 7.5 h. One group of animals was given 200 micrograms cycloheximide per 100 g body weight intramuscularly 1 h before start of the perfusion. Serial estimations of protein and of four brush border membrane enzymes (alanine aminopeptidase, alkaline phosphatase, gamma-glutamyl transferase, and enteropeptidase) were done in the perfusate. The results provide evidence that during the experiments an increasing proportion of the enzymes stems from de novo synthesis. This is consistent with the concept that after loss of 10-30 per cent of enzyme the molecules are replaced by newly synthesized material, provided that the energy metabolism of the mucosa cells remains intact.
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PMID:De novo synthesis of brush border membrane enzymes during intestinal perfusion with bile salt in the rat. 614 76

Porcine rotaviral infectivity for continuous porcine kidney (PK-15) cells was enhanced by incorporation of pancreatic endopeptidases into the cell culture maintenance medium. Marked enhancement of infectivity was induced by trypsin, whereas elestase and alpha-chymotrypsin enhanced infectivity to a lesser extent. Bacterial protease also induced some enhancement of porcine rotaviral infectivity. A synergistic enhancement of porcine rotaviral infectivity was noticed with trypsin and alpha-chymotrypsin combined. Porcine rotaviral infectivity was not affected by incorporation of alpha-amylase, alkaline phosphatase, beta-galactosidase, carboxypeptidase-A, deoxyribonuclease, enterokinase, lipase, or ribonuclease into the maintenance medium.
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PMID:Porcine rotaviral infection of cell culture: effects of certain enzymes. 624 64

Suckling mice infected with reovirus type 3 were examined for changes in the epithelial brush border of the small intestine. After 3 days of infection with reovirus type 3, no significant changes were found in intestinal morphology or activity of any enzymes tested. After 6 days, villi were shortened and blunted with lymphangiectatic lesions and mild mononuclear infiltration in the lamina propria. In addition, there was a significant decrease in lactase (P < 0.001) and enterokinase activity (P < 0.05). However, there were no significant changes in the activities of alkaline phosphatase. In contrast, maltase (P < 0.001) and leucine aminopeptidase (P < 0.05) activities in the infected mice were significantly increased. These data suggest that brush border enzymes are affected differently by reovirus infection.
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PMID:Small intestinal epithelial brush border enzymatic changes in suckling mice infected with reovirus type 3. 625 Jan 21

Bovine enterokinase was incorporated into vesicles reconstituted from a soybean phospholipid mixture. A thin film hydration procedure (MacDonald, R. I., and MacDonald, R. C. (1975) J. Biol. Chem. 250, 9206-9214) produced vesicles with 40% of the enterokinase activity bound in the membrane. The highest incorporation was observed when cholesterol or dimyristoylphosphatidylethanolamine was added to the soybean phospholipids. Crude and highly purified enterokinase preparations were incorporated to the same extent suggesting that other membrane components were not required for a successful reconstitution. The properties of enterokinase in phospholipid vesicles were compared with those of alkaline phosphatase, which was also added to the reconstitution system, and with the enzyme activities present in vesicles prepared from brush-border membranes. The enzyme activities were not released by solutions of high ionic strength and remained associated with the phospholipid vesicles on gel filtration, ultracentrifugation, and sucrose density centrifugation. Enterokinase and alkaline phosphatase had their active sites exposed to substrate in the brush-border membrane vesicles. In soybean phospholipid vesicles half of the active sites of both enzymes were on the outside, since release of the enzyme with Triton X-100 almost doubled the units of enzyme present. Incubation of the soybean phospholipid and brush-border membrane vesicles with papain released the exposed molecules of enterokinase. The released enzyme molecules were fully active but could not be reincorporated into phospholipid vesicles. This suggests that the structure imbedded in the lipid bilayer was essential for a successful reconstitution. We conclude that the reconstituted soybean phospholipid vesicles are a suitable membrane system for the further study of membrane-bound enterokinase.
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PMID:Incorporation of bovine enterokinase in reconstituted soybean phospholipid vesicles. 633 12

The activities of maltase, lactase, alkaline phosphatase and enterokinase were followed in the small intestine of rats during prenatal development. These enzymes were detectable only after the 17th day of gestation. Furthermore, each enzyme exhibited a different pattern of prenatal presence. Maltase activity appeared first (day 18), followed by lactase and alkaline phosphatase (day 19) and then enterokinase (day 20). Except for enterokinase, all of the enzymes attained a level of activity close to the newborn levels at the final day of gestation. Induced intrauterine growth retardation during the 3rd trimester led to a decrease in intestinal weight proportional to the reduction of body weight. These decrease in size of the small intestine was caused by a reduction in cell number rather than cell size. Induced intrauterine growth retardation also resulted in a selective reduction in the specific activities of lactase and alkaline phosphatase, but not of enterokinase and maltase. These results suggest that reduction in maternofetal blood flow in the 3rd trimester of gestation will cause a selective decrease in some brush border enzymes (lactase and alkaline phosphatase) but does not effect others (maltase and enterokinase).
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PMID:Effect of intrauterine growth retardation on the activities of fetal intestinal enzymes in rats. 678 32

Extracellular production of recombinant proteins in Escherichia coli has several advantages over cytoplasmic or periplasmic production. However, nonpathogenic laboratory strains of E. coli generally excrete only trace amounts of proteins into the culture medium under normal growth conditions. Here we report a systematic proteome-based approach for developing a system for high-level extracellular production of recombinant proteins in E. coli. First, we analyzed the extracellular proteome of an E. coli B strain, BL21(DE3), to identify naturally excreted proteins, assuming that these proteins may serve as potential fusion partners for the production of recombinant proteins in the medium. Next, overexpression and excretion studies were performed for the 20 selected fusion partners with molecular weights below 40 kDa. Twelve of them were found to allow fused proteins to excrete into the medium at considerable levels. The most efficient excreting fusion partner, OsmY, was used as a carrier protein to excrete heterologous proteins into the medium. E. coli alkaline phosphatase, Bacillus subtilis alpha-amylase, and human leptin used as model proteins could all be excreted into the medium at concentrations ranging from 5 to 64 mg/L during the flask cultivation. When only the signal peptide or the mature part of OsmY was used as a fusion partner, no such excretion was observed; this confirmed that these proteins were truly excreted rather than released by outer membrane leakage. The recombinant protein of interest could be recovered by cleaving off the fusion partner by enterokinase as demonstrated for alkaline phosphatase as an example. High cell density cultivation allowed production of these proteins to the levels of 250-700 mg/L in the culture medium, suggesting the good potential of this approach for the excretory production of recombinant proteins.
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PMID:Proteome-based identification of fusion partner for high-level extracellular production of recombinant proteins in Escherichia coli. 1872 29

Immunoassays are representative biochemical detection methods. Among them, sandwich-type immunoassays, typified by sandwich ELISA, have used in disease diagnosis or biochemical detection with high target selectivity. Horseradish peroxidase and alkaline phosphatase have been typically used for signal amplification in ELISA. Recently developed sandwich-type immunoassays such as biobarcode immunoassays, immuno-PCR, and immuno-RCA have improved sensitivity by changing mainly the signal amplification method. To develop a novel amplification method in ELISA, an enzyme-cascading system was incorporated into an ELISA, and the new assay is termed a cascading enzyme-linked immunosorbent assay (CELISA). This CELISA includes a trypsinogen-enterokinase combination as the cascading enzyme system, and was used to detect alpha-fetoprotein (AFP), which is a liver cancer marker, and prostate-specific antigen (PSA). Using a colorimetric reagent for signal generation, CELISA had 0.1-10pM limits-of-detection for AFP and PSA in whole human serum and assay buffers, depending on the platform, well plate, or microbead type used. This study represents the first example that incorporated an enzyme cascading step in an ELISA system, resulting in successful signal amplification with sensitive detection of pathogenic antigens in serum.
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PMID:Cascade enzyme-linked immunosorbent assay (CELISA). 1966 63


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