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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 rapid batch procedure is described for purification of T4 polynucleotide kinase (ATP:5'-dephosphopolynucleotide 5'-phosphotransferase, EC 2.7.1.78) to near homogeneity using Blue Dextran-Sepharose chromatography. The enzyme preparation is sufficiently free of contaminating endonuclease and alkaline phosphatase activities to be suitable for radioactively labeling nucleic acids in vitro. Kinetic measurements indicate that the chromophore of Blue Dextran, Cibacron Blue F3GA, inhibits the activity of T4 polynucleotide kinase competitively with respect to single stranded DNA substrate and non-competitively with respect to the rATP substrate.
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PMID:A rapid purification of T4 polynucleotide kinase using Blue Dextran-Sepharose chromatography. 21 25

An alkaline phosphatase (EC 3.1.3.1) of the placental type was isolated from a seminoma type of human testicular cancer tissue and was purified to homogeneity by sulfate-mediated chromatography on a column of Cibacron Blue Sepharose 4B. The purified enzyme had a specific activity of 40.6 kU per gram of protein and was obtained in a yield of 37%. The purification procedure used was simple and economical, and may be used to purify alkaline phosphatase isoenzymes from other cancer tissues. This is the first report of the purification of the enzyme in seminoma. Inhibition studies suggest that this enzyme is a Nagao variant rather than the Regan type reported in several cancer tissues.
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PMID:Isolation and purification of placental type alkaline phosphatase from a seminoma. 310 Jan 1

The interaction of alkaline phosphatase (EC 3.1.3.1) from calf intestine with different dyes, especially with Procion Red HE-3B was studied by several methods. From the kinetic analysis a nonlinear noncompetitive type of inhibition with an inhibition constant Ki = 0.03 mM for Procion Red HE-3B and Cibacron Blue F3G-A was estimated. The extent of inhibition of the two dyes at constant substrate and inhibitor concentration is 10 to 20 times higher than that of natural inhibitors like L-phenylalanine and NADH. Difference spectroscopic measurements with Procion Red HE-3B showed that the enzyme dimer possesses two binding sites for the dye. The dissociation constant of the dye-enzyme complex was estimated to be Kd = 0.01 mM. The binding of Procion Red HE-3B to the enzyme is mainly stabilized by electrostatic interactions. Large aromatic parts of a dye molecule like a combination of two naphthol ring systems or an anthraquinone ring flanked by spatially arranged charged substituents are important for the extent of specificity. The elution of the enzyme from the immobilized dye and the quenching of the dye-protein difference spectral signal by the competitive inhibitor phosphate and by substrates suggest the involvement of the active center of the enzyme in the dye binding region.
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PMID:Interaction of procion red HE-3B and other reactive dyes with alkaline phosphatase: a study by means of kinetic, difference spectroscopic and chromatographic methods. 344 94

A heme-controlled inhibitor of translation was isolated from the S-100 of rabbit reticulocytes by a novel procedure including chromatography on double-stranded ribonucleic acid (dsRNA)-cellulose. The inhibitor thus purified is extremely active and functionally resembles previously studied heme-controlled inhibitor preparations in terms of kinetics and extent of inhibition of translation, relief of inhibition by eukaryotic initiation factor 2 (eIF-2), relief of inhibition by 2-aminopurine, and preferential inhibition of alpha-over beta-globin synthesis. The action of this inhibitor on translation is resistant to treatment with bacterial alkaline phosphatase, micrococcal nuclease, or trypsin and to incubation at 95 degrees C, pH 2 or pH 12. The inhibitor not only is retained on DEAE-cellulose, phosphocellulose, and dsRNA-cellulose but also exhibits a high affinity for the dye Cibacron Blue, properties that suggest that it may be a protein. Unlike previously described heme-controlled inhibitor preparations, or preparations that did not pass over dsRNA-cellulose, the inhibitor recovered upon dsRNA-cellulose chromatography does not exhibit eIF-2 kinase activity. The inhibitor does not block ternary complex formation between eIF-2, methionyl-tRNAfMet, and GTP but inhibits the ability of eIF-2 to form a complex with labeled globin mRNA. In the presence of inhibitor, the formation of mRNA/eIF-2 complexes can be restored effectively by an excess of eIF-2 but not by an excess of mRNA. The inhibitor thus appears to block the interaction between eIF-2 and mRNA not by competing with eIF-2 for a binding site on mRNA but, instead, by acting on eIF-2 itself.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Isolation of a heme-controlled inhibitor of translation that blocks the interaction between messenger rna and eukaryotic initiation factor 2. 647 77

Possible interactions of human liver and intestinal alkaline phosphatases with Cibacron Blue F3GA were examined. The results indicated that the intestinal enzyme bound to the dye column whereas the liver enzyme did not. The affinity of intestinal alkaline phosphatase with the dye-ligand appeared to be biospecific, since a low concentration of purine nucleoside phosphates or potassium phosphate specifically reversed the binding. Taking advantage of the variant alkaline phosphatase from human hepatocellular cancer tissue to behave on the dye adsorbent in a similar fashion with the intestinal enzyme, it was purified by Cibacron Blue F3GA affinity chromatography, producing a 189-fold purification with a yield of 93%.
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PMID:Interaction of human intestinal and hepatoma alkaline phosphatases with immobilized Cibacron Blue F3GA. 683 May 99

Low concentrations of metal ions, particularly those of the first row transition series such as Zn2+, Co2+, Mn2+, Ni2+, Cu2+, and, to a lesser extent, the group IIA ions, Ca2+ and Mg2+, promotes binding of carboxypeptidase G2, alkaline phosphatase and yeast hexokinase to immobilized Procion Red H-8BN, Procion Yellow H-A and Cibacron Blue F3G-A respectively. The binding of ovalbumin to immobilized Cibacron Blue F3G-A and Procion Orange MX-G is selectively enhanced in the presence of AI3+. With ovalbumin and alkaline phosphatase, the effect is almost totally specific for both the metal ion and dye, whereas with carboxypeptidase G2 and hexokinase, metal ions such as Co2+, Ni2+, Mn2+, Cu2+, Ca2+ and Mg2+ also promote binding to varying degrees. Almost all other monovalent and trivalent metal ions appear to be ineffective. Metal ion-bound enzymes can subsequently be eluted with appropriate chelating agents of the amine, aminocarboxylate or substituted pyridine classes.
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PMID:Metal ion-promoted binding of proteins to immobilized triazine dye affinity adsorbents. 689 1

The paper deals with a simple and effective procedure for the isolation of calf intestinal alkaline phosphatase (EC 3.1.3.1) with a yield of 35 per cent by employing immobilized Procion Red HE-3B and Cibacron Blue F3G-A, respectively, as dye-ligands. The resulting enzyme is homogeneous and has a specific activity of about 2500 units per mg of protein. Because dye liganded gels are of low costs and can be used several times without loss of binding properties, the presented method is in particular suited for large scale application.
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PMID:Preparation of homogeneous alkaline phosphatase from calf intestine by dye-ligand chromatography. 710 Jan 27