Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:3.1.3.1 (alkaline phosphatase)
 47,916 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Two PRL-like glycoprotein hormone complexes were purified from the medium of cultured mouse conceptuses from day 10 of pregnancy: mouse placental lactogen-I (mPL-I) (29-32K), and mPL-I (36.5-42K). Sodium dodecyl sulfate-gel electrophoresis revealed that mPL-I (36.5-42K) is a complex of five proteins with mol wt of 36.5K, 37.5K, 39K, 40.5K, and 42K. Deglycosylation with peptide: N-glycosidase F or trifluoromethanesulfonic acid produced a single 29K protein. mPL-I (36.5-42K) was also sensitive to neuraminidase, but not to endo-beta-N-acetylglucosaminidase H or bacterial alkaline phosphatase. The production of intermediates from partial digestion of mPL-I (36.5-42K) with endo-beta-N-acetylglucosaminidase F indicated the presence of multiple glycosylation sites. mPL-I (29-32K) is a complex of three proteins with mol wt of 29K, 30.5K, and 32K. Treatment with peptide:N-glycosidase F or trifluoromethanesulfonic acid reduced the mol wt of the 30.5K and 32K bands to 28K. The 30.5K band was sensitive to endo-beta-N-acetylglucosaminidase H and endo-beta-N-acetylglucosaminidase F, but the 32K band was not. Neither band was sensitive to neuraminidase or bacterial alkaline phosphatase. The 29K band was resistant to all chemical and enzymatic treatments and is probably not glycosylated or phosphorylated. In the nonreduced state, neither form of mPL-I showed an increase in mobility over that of its reduced counterpart on sodium dodecyl sulfate-gel electrophoresis, indicating that neither form of mPL-I contains the large disulfide loop common to hormones of the PRL family. After iodination, all component proteins of both forms of mPL-I were found to bind to day 17 pregnant mouse liver membranes and were displaceable by excess mPL-II. In a radioreceptor assay, 125I-labeled mPL-I (36.5-42K) was displaced by mPRL or mPL-II, but not by mGH. An antiserum to both forms of mPL-I was generated, and a RIA employing mPL-I (36.5-42K) as the standard and radioligand was developed. Dilutions of day 10 pregnant maternal mouse serum and placental homogenate and a partially purified fraction of mPL-I (29-32K) produced displacement curves parallel to that of mPL-I (36.5-42K) standard curve. Five micrograms of mPRL, mPL-II, or mGH or 10 microliter day 17 pregnant or male mouse serum did not displace the radioligand from the antibody. mPL-I (36.5-42K) was lactogenic, but it did not possess LH-like bioactivity.
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PMID:Purification and partial characterization of two prolactin-like glycoprotein hormone complexes from the midpregnant mouse conceptus. 303 95

A modified agarose electrophoretic system for the separation of alkaline phosphatase (ALP, EC 3.1.3.1) isoenzymes is described. Bone, liver, high-molecular-mass, and intestinal ALP are separated with high reproducibility. The sensitivity of the agarose system is superior to cellulose acetate in detecting high-Mr ALP. Correlation is good between bone ALP fractions scanned before and after treatment with neuraminidase. Immunoglobulin-bound ALPs, the ALP-lipoprotein-X complex, and the additional ALP fraction observed in transient hyperphosphatasemia in children are detected by their peculiar electrophoretic mobility in the proposed system. Approximately 25% of the samples contained an additional fraction of intestinal-type ALP, as evidenced by neuraminidase treatment and use of polyclonal and monoclonal antibodies. Because the electrophoretic mobilities of this "intestinal variant" and of some immunoglobulin-bound ALP fractions are identical to those of bone and intestinal ALP, respectively, treatment of the samples with a polyclonal antibody that reacts with intestinal ALP is advised.
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PMID:Improved agarose electrophoretic method for separating alkaline phosphatase isoenzymes in serum. 304 80

A novel alkaline phosphatase (AP) isozyme was found in human adipose tissue. Adipose tissue alkaline phosphatase differed in enzymatic properties from liver, placental and intestinal alkaline phosphatases. On electrophoresis it showed the same mobility as intestinal alkaline phosphatase, but after treatment with neuraminidase its mobility was decreased to the same as or slightly less than that of neuraminidase-treated liver alkaline phosphatase. Its inhibition by amino acids, inactivation by urea and activation by Mg2+ were almost the same to those of liver alkaline phosphatase. However, at 56 and 65 degrees C it was more stable than liver alkaline phosphatase. Alkaline phosphatase activity was demonstrated histochemically in adipose tissue with naphthol AS-MX phosphate as substrate. It was localized in the wall of blood capillaries, but not present in adipocytes.
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PMID:A novel alkaline phosphatase isozyme in human adipose tissue. 310 Jan 9

This simplified HPLC method for measurement of high-molecular-mass alkaline phosphatase (high-Mr AP; EC 3.1.3.1) in serum and bile is rapid (time for column preparation and separation 30 min), reproducible (CV 4.2%), and highly sensitive (detects high-Mr AP in healthy controls at 1-3% of total AP activity in serum), and is suitable for processing small batches of sample. We characterized high-Mr AP in serum and bile by incubating samples with L-phenylalanine, neuraminidase, 1-butanol, or wheat-germ lectin, and by determining stability to heat. High-Mr AP activity was determined in sera of patients with various liver diseases (4-32% of total AP serum activity) and results were compared with those by electrophoresis on agarose.
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PMID:High-molecular-mass alkaline phosphatase: simplified and highly sensitive determination by liquid chromatography. 316 31

In five patients with benign transient hyperphosphatasaemia (THP), high activities of so-called "atypical" alkaline phosphatase or fragment isoenzymes were detected. One case occurred after rotavirus infection. Incubation with neuraminidase suggested that "atypical" alkaline phosphatase originated from highly glycosylated bone and liver isoenzymes. This may have been due to virus-induced low isoenzyme clearance from serum. The course of isoenzyme activities in THP following rotavirus infection was followed. Determination of atypical alkaline phosphatase may be useful in the diagnosis of THP.
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PMID:Transient hyperphosphatasaemia of infancy. 321 1

High-performance liquid chromatography using pellicular quaternary amine-bonded resins was used to separate a variety of neutral, sialylated, and phosphorylated oligosaccharides. At pH 4.6, sialylated compounds were separated according to number of negative charges, sialic acid linkage [alpha(2,3) compared to alpha(2,6)], and position of sialic acid linkage along a linear saccharide chain. At pH 13, the neutral sugar portion of the sialylated chain had a significant effect on the separation, due to oxyanion formation. Specifically, sialylated tetrasaccharides containing the Gal beta(1,3)GlcNAc sequence were retained much more than their Gal beta(1,4)GlcNAc- or Gal-beta(1,4)GalNAc-sialylated counterparts. Linear phosphorylated oligosaccharides could be completely separated according to number of charges and net carbohydrate content. Partial separation of linear-chain positional isomers, differing in either location of Man-6-PO4 in the chain or linkage position of Man or Man-6-PO4, was accomplished. Branched-chain phosphorylated compounds could be completely separated according to which antennae contained the Man-6-PO4. The electrochemical current generated by oxidation of sialylated, phosphorylated, and neutral oligosaccharides was compared to that of a glucose. The relative molar response factors for neutral, sialylated, and phosphorylated oligosaccharides ranged from 0.2 to 3.2. Neutral oligosaccharides gave the following molar responses for each group of structurally related compounds: (1) mono- and disaccharide, 1-1.3; (2) linear tri- and tetrasaccharides, 1.5-2.0; and (3) branched pentasaccharide-nonasaccharides, 2.4-3.1. Response factors for the sialyated compounds were not as consistent and were affected by linkage position of sialic acid. For oligosaccharides of the same size, increasing phosphorylation resulted in a twofold decrease in response factor for each added phosphate group. Therefore, conversion of sialylated and phosphorylated oligosaccharides to their neutral counterparts, using alkaline phosphatase or neuraminidase, respectively, was required for quantitative analysis of oligosaccharide mixtures using electrochemical response. Using this approach, complete separation of the parent neutral structures was obtained, the relative proportions of the neutral species were quantified, and the amount of sialic acid released was easily determined in a neuraminidase digest.
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PMID:High-performance anion-exchange chromatography of oligosaccharides using pellicular resins and pulsed amperometric detection. 323 49

A simple and new method has been developed to detect apolipoprotein E polymorphism directly from plasma or serum without prior ultracentrifugation and delipidation. The method combines the use of dialyzed plasma or serum samples with or without neuraminidase treatment followed by monodimensional isoelectric focusing in simple or 3 M urea gels at a constant low power and progressively increasing voltage over a 3-hr period, and finally protein blotting to a nitrocellulose membrane. Apolipoprotein E phenotypes are identified immunologically using a double antibody reaction, the primary antibody being a monospecific, polyclonal goat anti-apolipoprotein E, and the secondary antibody being a rabbit anti-goat IgG conjugated with alkaline phosphatase. The method was employed to screen apolipoprotein E polymorphism in two white populations in the United States. The frequency values are comparable to those reported previously by other investigators using conventional detection methods. The procedure is simple, accurate, suitable for large scale epidemiologic, clinical, and genetic studies.
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PMID:Genetic studies of human apolipoproteins. V. A novel rapid procedure to screen apolipoprotein E polymorphism. 324 Nov 28

Three acid phosphatase (EC 3.1.3.2) allozymes (ACPH1, ACPH2, and ACPH4) of Drosophila virilis show different activities as measured by electrophoretic techniques. Recently, it was suggested that these differences are attributable to the variable ability of the allozymes to be incorporated into lysosomes (Narise, S., Genet. Res. Cambr., 45:143, 1985). Immunoelectrophoresis demonstrated that the activity differences between these electrophoretic variants coincided with differences in the amount of the enzyme protein in soluble fractions but not in whole cell-free extracts. These results support the idea that acid phosphatase allozymes in D. virilis are cell-localization variants. We examined the problem by structural analysis of both the protein and the carbohydrate moieties of these allozyme glycoproteins, since lysosomal enzymes are known to become localized in lysosomes through their carbohydrate moieties. The three ACPH allozymes were purified to homogeneity from their respective homozygotes and compared with respect to amino acid composition and carbohydrate content and composition. Amino acid compositions were similar, while content and compositions of neutral sugars were significantly different. The neutral sugar content of ACPH1 was 9.2%; that of ACPH2, 21.0%; and that of ACPH4, 7.3%. A trace of hexosamines, but no N-acetylneuraminic acid, was found in the ACPH allozymes. Isoelectric points varied corresponding to their electrophoretic mobilities, which were not changed by treatment with alkaline phosphatase and neuraminidase.
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PMID:Comparison of purified acid phosphatase allozymes in Drosophila virilis: differences in carbohydrate content and composition of the allozymes. 330 79

In rat liver, alkaline phosphatase is shown to exist in several distinct molecular forms originating from different cell types. The alkaline phosphatases of isolated hepatocytes and non-parenchymal liver cells were characterized with respect to electrophoretic mobility, thermostability, and sensitivity to treatment with neuraminidase in order to define the cellular distribution of the different enzyme forms within the liver. The major form of liver alkaline phosphatase could be attributed to the hepatocytic enzyme, whereas the properties of the minor form were found to be identical with those of the non-parenchymal cell enzyme. In contrast to the hepatocytic enzyme, that of the non-parenchymal cells revealed heterogeneity after desialylation.
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PMID:Multiple forms of alkaline phosphatase in rat hepatocytes and non-parenchymal liver cells: differences in electrophoretic mobility, thermostability and sensitivity to neuraminidase treatment. 339 Jan 66

Washed formol-fixed normal human platelets have been separated into surface charge-dependent subpopulations using high voltage continuous flow electrophoresis. The procedure is highly reproducible and the heterogeneity profile extends over 20-25 fraction tubes on the anodal side of the entry port to the separation chamber. Fractions have been subdivided into subpopulation pools A, B and C which have mean mobilities by analytical cytopherometry extending over the range 0.81-0.91 micron/s/V/cm from the least (C) to the most (A) electronegative cells. Coulter volume differences across the profile from 5.0 to 12.8 fl correlated well with electrophoretic mobilities whereas buoyant density appeared to be an independent parameter. Analysis of surface neuraminidase-labile sialic acid of the platelets in pools A and C correlated well with differences in electrophoretic mobility, whereas a similar relationship for the alkaline phosphatase-labile phosphate moieties (also believed to be contributory to cell surface electrokinetic properties) could not be established even though in both cases the profiles of the enzyme-treated platelets showed significant shifts towards the cathode when compared with untreated cells. Titration of surface DTNB-reactive sulphydryl (-SH) groups revealed an inverse relationship between electronegativity and membrane -SH group status. This electrophoretic expression of subpopulation heterogeneity within the circulating platelet pool may have advantages in studying clinical conditions where the profiles may reflect cell surface interactions 'in vivo'.
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PMID:Platelet surface charge heterogeneity: characterization of human platelet subpopulations separated by high voltage continuous flow electrophoresis. 339 Mar 95


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