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)

A stable deoxyribonucleic acid (DNA) polymerase (EC 2.7.7.7) with a temperature optimum of 80 degrees C has been purified from the extreme thermophile Thermus aquaticus. The enzyme is free from phosphomonoesterase, phosphodiesterase and single-stranded exonuclease activities. Maximal activity of the enzyme requires all four deoxyribonucleotides and activated calf thymus DNA. An absolute requirement for divalent cation cofactor was satisfied by Mg2+ or to a lesser extent by Mn2+. Monovalent cations at concentrations as high as 0.1 M did not show a significant inhibitory effect. The pH optimum was 8.0 in tris(hydroxymethyl)aminomethane-hydrochloride buffer. The molecular weight of the enzyme was estimated by sucrose gradient centrifugation and gel filtrations on Sephadex G-100 to be approximately 63,000 to 68,000. The elevated temperature requirement, small size, and lack of nuclease activity distinguish this polymerase from the DNA polymerase of Escherichia coli.
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PMID:Deoxyribonucleic acid polymerase from the extreme thermophile Thermus aquaticus. 0 32

1. Alkaline phosphatase (orthophosphoric-monoester phosphohydrolase (alkaline optimum), EC 3.1.3.1) in guinea pig thymus was extracted optimally in 10 mM Tris - HCl buffer at pH 8.0 containing 5 g/l Triton X-100. 2. alpha-Glycerophosphate, beta-glycerophosphate and phenolphthalein monophosphate were hydrolyzed by thymus extract with a pH optimum at 9.8-10.0, whereas p-nitrophenylphosphate and alpha-naphthylphosphate were hydrolyzed with pH optima at 10.7-10.8 and beta-naphthylphosphate at pH 11.2. P-Nitrophenylphosphate and phenolphthalein monophosphate proved to be the most suitable substrates. 3. Alkaline phosphatase was effectively inhibited by EDTA, Zn2+, histidine and urea therefore resembling the inhibition characteristics of alkaline phosphatase in the placenta and kidney, but not that in the liver and intestine, which differed markedly. 4. DEAE-cellulose chromatography and polyacrylamide disc electrophoresis revealed three enzyme peaks which did not differ in their substrate specificities and modifier characteristics. 5. Polyacrylamide disc electrophoresis of thymus, serum, placenta, kidney, liver, bone and intestine revealed no alkaline phosphatase bands definitely unique to thymus.
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PMID:Biochemical characterization of alkaline phosphatase in guinea pig thymus. 1 86

2-Deoxy-D-galactose, in a dose of 3 mmol/kg, was administered intraperitoneally twice daily to young rats for periods up to 12 weeks. This dosage schedule resulted in recurrent phosphate trapping predominantly in liver. UTP deficiency was excluded by simultaneous uridine injections. Phosphate trapping was caused by the rapid accumulation of 2-deoxy-D-galactose 1-phosphate and was most pronounced in liver but also demonstrated in small intestine, brain, spleen, and thymus. The marked, although transient, drop in the hepatic content of inorganic phosphate triggered the catabolism of adenine nucleotides and a loss of ATP. Other metabolic pathways affected by phosphate deficiency include glycogenolysis and glycolysis. Increasing with time, repeated doses of the galactose analog led to retardation and arrest of growth, hepatomegaly, and splenomegaly. The average relative liver and spleen weights were elevated 2.5- and 4.5-fold, respectively, after 12 weeks of treatment. Liver damage was indicated by hyperbilirubinaemia and a progressive rise in the activity in plasma of sorbitol dehydrogenase, alkaline phosphatase, and gamma-glutamyltransferase. Examination by light and electron microscopy showed increasing numbers of vacuoles, surrounded by a single membrane, in hepatocytes, sinusoidal endothelial cells, and Kupffer cells. Focal cytoplasmic degeneration in hepatocytes was occasionally indicated by formation of autophagic vacuoles and finger print lysosomes. Hepatocytes of 2-deoxy-D-galactose-treated rats showed a dissociation and fragmentation of the rough endoplasmic reticulum. Sinusoidal endothelial cells and Kupffer cells were markedly enlarged, the latter contained a PAS-positive but amylase resistant substance. Extrahepatic changes included an increased occurrence of vacuolated cells in thymus. Phosphate trapping and its metabolic consequences are common phenomena in the experimental injury induced b 2-deoxy-D-galactose and in some hereditary diseases such as uridylyltransferase deficiency galactosaemia, fructose intolerance and glucose-6-phosphatase deficiency.
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PMID:Consequences of recurrent phosphate trapping induced by repeated injections of 2-deoxy-D-galactose. Biochemical and morphological studies in rats. 4 10

An antigenic substance reactive with autoantibodies found in patients with cancer and autoimmune diseases was isolated from calf thymus. The purification procedure included extraction of the tissues with acetone powder, batch and column chromatography on DEAE-resins, ammonium sulfate precipitation, gel filtration on Sephadex G-200, and affinity chromatography on antibody-Sepharose 4B. Indirect immunofluorescence examination of cultured human embryo cells, using the serum of patients with nasopharyngeal cancer, showed a speckled nuclear pattern. The antigenic factor was a soluble acidic protein with a pI of 5.0 and a molecular weight of 250,000. The antigenic activities of this purified substance from calf thymus, and of the material on the cultured human embryo cells, were destroyed by proteases, ribonuclease, and alkaline phosphatase. The determinants were also sensitive to periodate oxidation. Thermal stability to 60 degree C and pH stability between 2.6 and 8.5 were demonstrated. Cross-reactivity of the antigenic substance with antibodies isolated from individuals with cancer and autoimmune diseases was shown by immunofluorescence, with appropriate blocking and absorption controls.
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PMID:Isolation of "speckled" nuclear antigen reactive with autoantibodies in patients with cancer and autoimmune diseases. 5 88

Monolayer and suspension cell cultures prepared from Hodgkin's disease tumors in the spleen were examined microscopically and by cytogenetics, tested for lymphocyte and monocyte cell surface properties, and assayed for enzymes by histochemical and spectrophotometric techniques. Hodgkin's disease monolayer cultures were composed of rapidly proliferating round and polygonal cells that were capable of propagation in vitro for an indefinite period of time. Abnormal aneuploid chromosomes were found in short-term Hodgkin's disease monolayers that had been passaged 16-20 times, and in established cell lines carried in culture longer than 3 yr and passaged more than 200 times. Cells fromHodgkin's disease monolayers contained lysozyme (muramidase), fluoride-resistant alpha naphthol acetate esterase, acid and alkaline phosphatase, and chymotrypsin-like activity. The monolayers did not exhibit specific cell surface markers or phagocytosis. Suspension cultures derived from Hodgkin's disease monolayers were composed of cells with aneuploid karyotypes and similar enzymes. The Hodgkin's disease suspension culture cells had surface receptors for complement and IgGFc, lacked surface or cytoplasmic immunoglobulin, and did not form Erosettes, react with an antithymocyte serum, nor exhibit phagocytosis. Normal monolayer culture cells, derived from adult spleen and human fetal spleen and thymus, were composed of spindle cells with a diploid number of chromosomes that could be carried for only a finite period of time in vitro. Normal cultured cells contained similar esterases and phosphatases, but were devoid of lysozyme and chymotrypsin-like activity. The morphologic, cytogenetic, cell surface, and enzymatic findings indicate that our Hodgkin's disease monolayer and suspension cultures are composed of cells with many properties suggesting an origin from monocytes (macrophages) rather than lymphocytes or fibroblasts. The presence of aneuploid karyotypes is consistent with a neoplastic origin and derivation from a malignant cell of Hodgkin's disease.
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PMID:Tissue culture studies in Hodgkin's disease: Morphologic, cytogenetic, cell surface, and enzymatic properties of cultures derived from splenic tumors. 6 93

Small oligonucleotides from DNA and RNA have been separated according to their base composition by high-performance anion-exchange liquid chromatography on Partisil-10 SAX using triethylammonium acetate buffer as the eluent. Fifteen of the 16 possible deoxydinucleoside monophosphates and all 16 dinucleoside monophosphates have been separated. All pairs of sequence isomers were all resolved. The 15 commercially available deoxydinucleotides were resolved into 13 fractions. A good resolution of deoxytrinucleoside diphosphates isolated from an alkaline phosphatase-Mg2+-activated DNase I digest of calf thymus DNA was achieved by this technique. A large number of sequence isomers could be fully separated. The base sequence of the eluted individual constituents has been determined by their hydrolysis with snake venom and spleen phosphodiesterase followed by high-performance liquid chromatographic analysis of the nucleotides released. The eight trinucleoside diphosphates isolated from an alkaline phosphatase-pancreatic RNase digest of yeast RNA have also been separated according to base composition. Their sequence was determined as above. The described technique is fast and gave very good separation. Most of the sequence isomers could be separated. Moreover, the eluent triethylammonium acetate can easily be removed from column effluents by freeze-drying in order to facilitate subsequent sequence analysis of the eluted compounds. The observed elution orders of the sequence isomers obey certain rules which are discussed in detail.
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PMID:Separation of small DNA and RNA oligonucleotides by high-performance anion-exchange liquid chromatography. 9 13

The kinetics of alkaline phosphatase active cells is followed up by quantitation in the thymus of newborn rabbits, of 1, 2, 3, 4, and 6 months old, as well as in adult ones. The proportion of these cells appears to be the lowest in newborn rabbits (2 per cent), increases up to 8 per cent in the 2-month-old and up to 15.3 per cent in the 3-month old rabbits. This maximal proportion rapidly decreases one month later, reaching values round about 1 per cent of all thymus cells in adult rabbits. This kinetics seems to reflect the process of functional maturation of the rabbit lymphoid system, the alkaline phosphatase active cells representing the thymocytes which differentiate in T cells or a subpopulation of them only.
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PMID:Kinetics of alkaline phosphatase active cells in the thymus of growing rabbits. 13 71

Novoimanine is an antibacterial drug from Hypericum perforatum L. When used in the bacteriostatic concentration, i.e. 0.5 gamma/ml, it induced release of potassium ions from the cells of Staphylococcus aureus 209P and had no effect on release of the UV-absorbing compounds and 14C-amino acids. In addition, incubation of the cells with novoimanine (2.5--50 gamma/ml) provided "preservation" in them of the earlier absorbed 14C-amino acids, while in the control cells their level decreased. In a concentration of 100 gamma/ml novoimanine stimulated activity of ATP-ase and alkaline phosphatase by 34 and 37-57 per cent respectively. Histones F1 and F3 of the calf thymus induced an intensive release of 14C-amino acids from the cells of staphylococci and increased the activity of ATP-ase by 6-10 times. The data of the study suggested that the effect of novoimanine on the cytoplasmic membrane was limited and different from that on the polycationic antibacterial agents.
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PMID:[Effect of novoimanine on the cellular permeability indices of staphylococci]. 14 41

Results on morphologic and hematologic characterization of a hamster leukemia capable of both cellular and cellfree transmission are described. Solid tumors removed in the course of several passages of leukemia animals, after producing blood smears, and spleen, liver, lymph nodes, bone marrow, thymus, kidney and lung were investigated histologically and histochemically. The morphological picture of the hamster leukemia has not changed during several transplantation generations. In addition to solid tumours, typical leukemic infiltration were detected histologically in spleen, liver, lymph nodes, bone marrow, kidneys and lung. No leukemic proliferation was noticed in the thymus. The final stage of the disease is characterized by an abrupt occurrence of high leukemic cell counts. The demonstration of alkaline phosphatase, but especially of naphtol-AS-D-chloroacetate esterase, in the leukemic cells is interpreted as indicating malignization of cells of the granulocytic line.
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PMID:[A transplantable myeloid hamster leukemia with high peripheral leukocyte counts and C-particles. II. Histology and cytochemistry (author's transl)]. 17 Aug 82

Studies on the maturational lineages of thymic lymphocytes have revealed several subclasses which are distinguishable on the basis of cell size, topographic distribution within the thymus, DNA synthetic and mitotic activity, migratory behavior, and other properties. Strain C57BL/Ka mice were inoculated with radiation leukemia virus at different concentrations, and tissues were removed at defined intervals. Sequential sections were analyzed for virus-specific cytoplasmic antigen expression, for morphological evidence of neoplastic transformation, and for alkaline phosphatase activity. The first detectable sign of MuLV infection was the focal appearance of cytoplasmic viral antigens in cells of the outer thymic cortex, followed by coalescence of such foci and, several weeks later, by the appearance of morphologically transformed and alkaline phosphatase-positive cells, again often focally distributed in the outer thymic cortex. These observations strongly suggest that the large, mitotically active cells of the outer thymic cortex are the principal source of target cells for both productive infection and subsequent lymphoma induction by the virus.
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PMID:Focal infection and transformation in situ of thymus cell subclasses by a thymotropic murine leukemia virus. 17 27


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