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:3.1.3.1 (alkaline phosphatase)
47,916 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Linear and branched glycopeptides containing multiple sialyl-N-acetyllactosamine side chains have been synthesized using a combined chemical and enzymatic approach. Peptide backbones in which beta-GlcNAc-Asn residues were incorporated were obtained in good yields by optimized solid-phase synthesis following the Boc strategy. The resulting multivalent glycopeptides were galactosylated in near-quantitative yields using bovine galactosyltransferase, UDP-galactose, and calf alkaline phosphatase that destroys the inhibiting side product UDP. Subsequent enzymatic sialylation yielded the desired glycopeptides containing asparagine-linked sialyl-N-acetyllactosamine side chains. The compounds were characterized by 1H NMR and FABMS. Recombinant sialyltransferase and CMP-sialate synthetase were used for the enzymatic synthesis of sialosides on a preparative scale. The synthetic glycopeptides were tested as inhibitors of influenza virus to cells, revealing that most of the multivalent sialoglycopeptides exhibit increased binding that depends on the spacing when compared to monovalent compounds. A possible mechanism for increased binding is proposed.
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PMID:Chemical and enzymatic synthesis of multivalent sialoglycopeptides. 814 76

Studies were performed on 670 male mice of Swiss strain. Cernilton (mixture of preparation Cernitin T-60-100 mg/kg and Cernitin GBX--20 mg/kg emulsified by using Imwitor 370) was intraperitoneally administered in a volume of 30 ml/kg of body mass. LD50 of paracetamol was fixed after intraperitoneal administration. Certimilton was given 1 h before or 1 h after paracetamol in a dose LD100 and LD50, thereafter the survival time and the number of deceased animals were determined. The effect of Cernilton preparation on the lesion of the liver induced by paracetamol was studied in 5 groups with 10 mice each: group 1--control; group 2--paracetamol; group 3--Cernilton, after 1 h paracetamol; group 4--paracetamol; after 1 h Cernilton; group 5--Cernilton. Paracetamol was injected in the following manner: a--300 mg/kg in a single dose, estimation after 3 h, b--300 mg/kg in single dose--section after 24 h, c--230 mg/kg/24 h single dose--estimation after 24 h, d--230 mg/kg/24 h four times--section after 24 h, e--230 mg/kg/24 h--seven times--estimation after 24 h. The degree of hepatic lesion was evaluated on the basis of the activity of alanine and asparagine aminotransferase as well as alkaline phosphatase, total bilirubin concentration in serum, the content of reduced glutathione and cytochrome P-450 in the liver as well as histological and histochemical examinations (glycogen, lipids) of the liver. It has been disclosed that Cernilton increases the survival rate of animals and decreases the hepatic lesion in the course of acute paracetamol intoxication, Cernilton is the factor that effectively normalizes the biochemical and morphological indices of hepatic lesions having been caused by repeated use of toxic paracetamol dose. The therapeutic action of pollen extracts is more effective than prophylactic one. The role of glutathione is significant in the mechanism of protective activity of the pollen extract.
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PMID:[Experimental evaluation of the effect of pollen extract on the course of paracetamol poisoning]. 815 23

Seven patients with primary biliary cirrhosis were treated with rifampicin administered for 2 weeks in a daily dose 450-600 mg. Due to the treatment the itch disappeared completely in 4 and decreased significantly in 3 patients. As shown by the antipyrine test, half-life and clearance of antipyrine returned to normal suggesting cytochrome P-450 induction as a result of hydroxylation activity. There was a tendency to lowering of bilirubin, cholesterol, alkaline phosphatase, asparagine--and alanine aminotransferase against an increase in gammaglobulins. The differences were, however, insignificant. Rifampicin tolerance was satisfactory.
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PMID:[The treatment of primary biliary liver cirrhosis with rifampicin]. 821 5

We have developed a method for monitoring the N-glycosylation of recombinant glycoproteins directly from conditioned medium samples. Proteins in the conditioned medium are separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and electroblotted onto polyvinylidene fluoride membranes. After staining the membranes with Coomassie blue, the protein(s) of interest is excised. Oligosaccharides are released from the membrane-bound glycoprotein by digesting with peptide N4-(acetyl-beta-glucosaminyl) asparagine amidase and labeled with the fluorophore 8-aminonaphthalene-1,3,6-trisulfonate (ANTS). Labeled oligosaccharides are then separated on polyacrylamide gels which allow for the direct comparison of samples. We have shown that recombinant human lysosomal hydrolase alpha-galactosidase A is N-glycosylated with both sialylated and phosphorylated oligosaccharides. ANTS-labeled oligosaccharide bands from alpha-galactosidase A were isolated from polyacrylamide gels. Sialylated and phosphorylated bands were identified by shifts in their electrophoretic mobility after digesting with neuraminidase or alkaline phosphatase to remove sialic acid or phosphate groups, respectively. Using the ANTS-labeled oligosaccharides from alpha-galactosidase A, we have shown that polyacrylamide gels can be used to resolve sialylated and phosphorylated oligosaccharide structures.
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PMID:A method for monitoring the glycosylation of recombinant glycoproteins from conditioned medium, using fluorophore-assisted carbohydrate electrophoresis. 857 98

In each subunit of the homodimeric enzyme Escherichia coli alkaline phosphatase, two of the three metal cofactors Zn2+ and Mg2+, are bound by an aspartate side-chain at position 51. Using site-specific mutagenesis, Asp51 was mutated both to alanine and to asparagine to produce the D51A and D51N enzymes, respectively. Over the range of pH values examined, the D51A enzyme did not catalyze phosphate ester hydrolysis above non-enzymic levels and was not activated by the addition of millimolar excess Zn2+ or Mg2+. Replacement of Asp51 by asparagine, however, resulted in a mutant enzyme with reduced activity and a higher pH optimum, compared with the wild-type enzyme. At pH 8.0 the D51N enzyme showed about 1% of the activity of the wild-type enzyme, and as the pH was raised to 9.2, the activity of the D51N enzyme increased to about 10% of the value for the wild-type enzyme. Upon the addition of excess Mg2+ at pH 9.2, the D51N enzyme was activated in a time-dependent fashion to nearly the same level as the wild-type enzyme. The affinity for phosphate of the D51N enzyme decreased tenfold as the concentration of Mg2+ increased. Under optimal conditions, the k(cat)/K(m) ratio for the D51N enzyme indicated that it was 87% as efficient as the wild-type enzyme. To investigate the molecular basis for the observed kinetic differences, X-ray data were collected for the D51N enzyme to 2.3 angstroms resolution at pH 7.5, and then to 2.1 angstroms resolution at pH 9.2 with 20 mM MgCl2. The two structures were then refined. The low magnesium, low pH D51N structure showed that the third metal site was unoccupied, apparently blocked by the amide group of Asn51. At this pH the phosphate anion was bound via one oxygen atom, between the zinc cations at the first and second metal sites, which strongly resembled the arrangement previously determined for the D153H enzyme at pH 7.5. In the high magnesium, high pH D51N structure, the third metal site was also vacant, but the phosphate anion bound closer to the surface of the enzyme, coordinated to the first metal site alone. Electron density difference maps provide evidence that magnesium activates the D51N enzyme by replacing zinc at the second metal site.
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PMID:Kinetic and structural consequences of replacing the aspartate bridge by asparagine in the catalytic metal triad of Escherichia coli alkaline phosphatase. 864 34

Out of 12 patients with primary hyperlipidemia (HL) included in the study ischemic heart disease was diagnosed in 4 and hypertension stage I-II in 6 patients. HL stage II B, III, IV and V was registered in 7, 2, 2 and 1 patients, respectively. Hypolipidemic therapy included hypolipidemic diet and Lipanor (Sanofi-Chinoin-Winthrop, France) in a dose 100 mg once a day after evening meal. The course lasted 3 months. Blood serum was examined for concentrations of total cholesterol (CS), triglycerides (TG), CS of HDLP, glucose, uric acid, prothrombin, fibrinogen, alanine and asparagine transaminase, alkaline phosphatase, creatinine, total bilirubin. After 1 month of lipanol treatment mean total CS concentration in the serum fell by 21.9% and remained such for 2-3 months of treatment. 1 month after the drug discontinuation mean concentration of total CS was under the initial one by 4.65%. Mean serum concentrations of TG 1 month after treatment decreased by 41.7%, after 2 months by 48.8%. Mean concentration of HDLP CS after 1 month of treatment rose by 19.2% and by 33.1% after one more month. 1 month after Lipanor discontinuation mean HDLP CS concentration exceeded the baseline level by 16%. Fibrinogen and prothrombin index declined. Hypolipidemic effect of Lipanor varied from case to case.
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PMID:[A trial of the use of the hypolipidemic preparation Lipanor (ciprofibrate) in patients with primary hyperlipidemia]. 877 89

A partial structure of many glycoproteins, a glycosylated asparagine carrying a complex type undecasaccharide N-glycan (Neu5Ac(alpha 2-6)Gal(beta 1-4)GlcNAc(beta 1-2)Man alpha 1-3) [Neu5Ac(alpha 2-6)Gal(beta 1-4)GlcNAc(beta 1-2)Man(alpha 1-6)]Man(beta 1-4) GlcNAc(beta 1-4)GlcNAc-Asn) was obtained by total synthesis. As a starting material served a chemically synthesized diantennary heptasaccharide azide which was deprotected in a three-step sequence in high yield. The reduction of the anomeric azide was accomplished with propanedithiol in methanol-ethyldiisopropylamine. Coupling of the glycosyl amine to an activated aspartic acid gave the benzyl protected asparagine conjugate. After removal of the six benzyl functions the resulting free heptasaccharide asparagine was elongated enzymatically in the oligosaccharide part. The use of beta-1,4-galactosyltransferase and alpha-2,6-sialytransferase in the presence of alkaline phosphatase allowed the efficient transfer of four sugar units to the acceptor resulting in a full length N-glycan, a sialyated diantennary undecasaccharide-asparagine of the complex type.
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PMID:Chemoenzymatic synthesis of a sialylated diantennary N-glycan linked to asparagine. 964 61

The BTI-Tn5B1-4 insect cell line, commercially available as the High Five cell line (Invitrogen), supports higher levels of recombinant protein production compared to existing insect cell lines. Proprietary serum-free media such as ExCell 405 (JRH Biosciences), Express Five (Life Technologies), IS BAC (Irvine Scientific), and CCM3 (HyClone) are available which were developed specifically for a suspension culture of High Five cells. While these media are highly optimized, a lower cost alternative is desirable for large-scale protein production which is also serum-free and supports good cell growth (>5 x 10(6) cells/mL) and recombinant protein production (>50 mg/L of secreted protein). The amino acid and carbohydrate metabolism of the Tn5B1-4 cells was first examined. It was found that asparagine was nearly depleted during batch growth in Ex-Cell 405, without limitations in glutamine, other amino acids, or glucose. Alanine also accumulated to about 35 mM during growth. We then extended the formulation techniques for medium development used for Spodoptera cell lines to the Tn5B1-4 cell line. A medium based on IPL-41 basal medium, Hy-Soy protein hydrolysate (Quest, International), yeastolate ultrafiltrate, a lipid-sterol emulsion, and Pluronic F-68 was developed. Dextran sulfate (100 microg/mL) was used to induce a single cell suspension culture. This medium is denoted as ISYL and performs best when supplemented with a 2.5% lipid-Pluronic F-68 mixture. Supplementation with additional aspargine in a 1.5% lipid-Pluronic F-68 mixture did not improve growth, suggesting that a lipid was growth-limiting and not an amino acid. Ex-Cell 405 and ISYL with 2.5% lipid-Pluronic F-68 supplement supported virtually identical growth rates, extent of growth (ca. 6.0 x 10(6) cells/mL) in an 80% oxygen atmosphere, and supported production of SEAP (secreted human alkaline phosphatase) at a volumetric level of about 65-70 mg/L. Thus, the less expensive ISYL medium can deliver acceptable performance and may be suitable for large-scale insect cell cultures.
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PMID:Low-cost serum-free medium for the BTI-Tn5B1-4 insect cell line. 969 78

The efficacy and safety of lomefloxacin in the treatment of patients with hepatitis due to the use of routine antituberculosis agents were estimated. The trial group included 20 patients (10 with increased activity of enzymes such as alanine and asparagine transaminases, alkaline phosphatase and gamma-glutamate dehydrogenase) who were treated for various forms of tuberculosis with antitubeculosis drugs. The treatment course with lomefloxacin was 4 weeks (400 mg twice a day at 12-hour intervals). The criteria of the enrolment to the trial group were a more than 2-3 times higher activity of the enzymes and the absence of the markers of the virus hepatitis A, B and C. The therapy efficacy before and after the use of lomefloxacin was estimated clinically and by the findings of the laboratory and instrumental investigations. As a result of the treatment with lomefloxacin normalization of the enzyme activity and a favourable time course of the main disease were observed in the patients with drug hepatitis due to the use of antituberculosis agents requiring continuation of the antituberculosis therapy. An important result of the complex treatment with lomefloxacin and antituberculosis agents was discontinuation of the tubercle bacilli isolation in 70 per cent of the patients. Lomefloxacin proved to be a safe and efficient up-to-date agent for the treatment of tuberculosis in patients with hepatitis due to the use of antituberculosis drugs.
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PMID:[Lomefloxacin in phthisiatric practice]. 982 4

A clinical observation over 57 liquidators of the Chernobyl accident aftermath (ChAAL) with associated diseases of the digestive and hepato-biliary system revealed abnormalities in blood enzymic activity, presenting as elevation of, in particular, alanine aminotransferase as well as of asparagine aminotransferase, gamma-glutaminetranspeptidase, alkaline phosphatase, lactate dehydrogenase, and of blood mineral composition at the expense of an increase in certain minerals. Disturbances in underlying process of bodily metabolism promote the development and association of erosive and ulcerous lesions of the stomach and duodenum with chronic noncalculous cholecystitis, persistent hepatitis in ChAAL.
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PMID:[Metabolic disorders in digestive and hepatobiliary pathology in those who worked in the cleanup of the aftermath of the accident at the Chernobyl Atomic Electric Power Station]. 1005 Apr 54


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