Different lectin affinities in rat alkaline phosphatase isozymes: Multiple forms of the isozyme isolated by heterogeneities of sugar moieties

Koyama, Isamu; Sakagishi, Yoshikatsu; Komoda, Tsugikazu

doi:10.1016/s0378-4347(00)83252-1

Cited by 39 publications

(17 citation statements)

References 22 publications

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“…For instance, he found that AP-ase isolated from duodenal mucosal cell homogenate eluted from Con A Sepharose into three frac tions: unbound, weakly bound and strongly bound. The relative content of the three fractions in duodenal homog enate was 46,48 and 6% for unbound, weakly bound and strongly bound fractions, respectively [20], We have ex tended this findings by showing recently that the same technique can be used to distinguish between the soluble and membranous isoenzymes of duodenal AP-ase [13]. The unbound fraction of AP-ase was dominant (65%) in the membranous AP-ase [13].…”

Section: Discussionmentioning

confidence: 95%

“…Con A chromatography was used to iso late AP-ase from different organs [20], Koyoma et al [20] found that the elution profile of AP-ase from a Con A col umn was different in different organs. For instance, he found that AP-ase isolated from duodenal mucosal cell homogenate eluted from Con A Sepharose into three frac tions: unbound, weakly bound and strongly bound.…”

Section: Discussionmentioning

confidence: 99%

“…The Con A elution profile diversity of AP-ase can be attributed to sugar chain heterogeneity [20,21]. Many authors have shown that the unbound fraction I has multiantennary or bisecting complex-type sugar chains; the weakly bound fraction II has biantennary complex-type sugar chains and the strongly bound fraction III has hybrid-or high-mannose-type sugar chains.…”

Section: Discussionmentioning

confidence: 99%

“…However, the fraction III seems to be very important as an indicator of the meta bolic activity of the cells. The low level of fraction III increased markedly in duodenal explants by the addition of swainsonine, an inhibitor of lysosomal and Golgi a-Dmannosidase II, and in liver cells after administration of cholera toxin or after bile duct ligation [20]. Furthermore, in the serum of streptozotocin-induced diabetic rats, the content of fraction III was also increased [23].…”

Section: Discussionmentioning

confidence: 99%

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Abnormal Glycosylation of Duodenal Membranous Alkaline Phosphatase Induced by Cysteamine in Rats

Japundzić

Kosaric²

1994

Digestion

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The membranous and soluble isoenzymes of alkaline phosphatase (AP-ase) were isolated from the duodenal mucosal cells in control and cysteamine-treated rats. Both the cytosolic and membranous isoenzymes of the AP-ase were drastically inhibited by a single subcutaneous injection of cysteamine-HCl, a potent duodenal ulcerogen. However, cysteamine did not change the isoenzyme distribution between the two compartments of the duodenal mucosal cells. Membranous isoenzyme of duodenal AP-ase in control rats was separated by concanavalin A (Con A) affinity chromatography into three fractions: unbound (65%), weakly bound (11 %) and strongly bound (24%). The strongly bound fraction disappeared completely from duodenal mucosal cells of cysteamine-treated rats before any visible signs of ulcer formation. Since the strongly bound fraction of AP-ase contains high mannose-type sugar chains, we concluded that the ulcerogen affected the initial mannosylation (or glypiation) of membranous AP-ase which took place in the rough endoplasmic reticulum of the mucosal cells. In control rats, the Con A fractions of the membranous AP-ase were separated by gel filtration as tetramers of M_r 420 and 340 kD and dimers of M_r 185-200 kD. In cysteamine-treated rats, neither the strongly bound tetramers of 340 kD nor dimers of 185 kD were found by gel filtration. However, cysteamine did not change the total tetramer:dimer ratio. Judging by the decrease of V_max values at the pH optimum, the different molecular forms of the AP-ase are selectively sensitive to cysteamine inhibition. The kinetic data show that the type of the tissue-specific inhibition by cysteamine is different for dimeric and tetrameric forms of the membranous AP-ase. It is of an ‘uncompetitive’ type for a tetrameric form and of a ‘non-competitive’ type for dimeric forms.

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Section: Discussionmentioning

confidence: 95%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

See 2 more Smart Citations

Abnormal Glycosylation of Duodenal Membranous Alkaline Phosphatase Induced by Cysteamine in Rats

Japundzić

Kosaric²

1994

Digestion

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“…Both the concanavalin A (Canavalia ensiformis, ConA)-Sepharose 4B (Sigma-Aldrich, MO) and the wheat germ agglutinin (Triticum vulgaris, WGA)-agarose CL-4B (Fluka, SG, Switzerland) columns were equilibrated with TBS (10 mM Tris-HCl, pH 8.0, supplemented with 0.5 M sodium chloride, 1 mM calcium chloride, 1 mM magnesium chloride, 1 mM manganese chloride, and 0.01 mM zinc chloride) at a flow rate of 0.2 ml/min. Lectin affinity chromatography was performed as described previously [47]. Briefly, the purified enzyme in 0.6 ml of TBS was applied to the ConA and WGA columns, and left to stand for 3h at room temperature.…”

Section: Lectin Affinity Chromatographymentioning

confidence: 99%

Enhancement of drug delivery to bone: Characterization of human tissue-nonspecific alkaline phosphatase tagged with an acidic oligopeptide

Nishioka

Tomatsu

Gutiérrez

et al. 2006

Molecular Genetics and Metabolism

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Hypophosphatasia is caused by deficiency of activity of the tissue-nonspecific alkaline phosphatase (TNSALP), resulting in a defect of bone mineralization. Enzyme replacement therapy (ERT) with partially purified plasma enzyme was attempted but with little clinical improvement. Attaining clinical effectiveness with ERT for hypophosphatasia may require delivering functional TNSALP enzyme to bone. We tagged the C-terminal-anchorless TNSALP enzyme with an acidic oligopeptide (a six or eight residue stretch of L-Asp), and compared the biochemical properties of the purified tagged and untagged enzymes derived from Chinese hamster ovary cell lines. The specific activities of the purified enzymes tagged with the acidic oligopeptide were the same as the untagged enzyme. In vitro affinity experiments showed the tagged enzymes had 30-fold higher affinity for hydroxyapatite than the untagged enzyme. Lectin affinity chromatography for carbohydrate structure showed little difference among the three enzymes. Biodistribution pattern from single infusion of the fluorescence-labeled enzymes into mice showed delayed clearance from the plasma up to 18h post infusion and the amount of tagged enzyme retained in bone was 4-fold greater than that of the untagged enzyme. In vitro mineralization assays with the bone marrow from a hypophosphatasia patient using each of the three enzymes in the presence of high concentrations of pyrophosphate provided evidence of bone mineralization. These results show the anchorless enzymes tagged with an acidic oligopeptide are delivered efficiently to bone and function bioactively in bone mineralization, at least in vitro. They suggest potential advantages for use of these tagged enzymes in ERT for hypophosphatasia, which should be explored.

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The mechanism of placental alkaline phosphatase induction in vitro

Nozawa

Narisawa

Iizuka

et al. 1989

Cell Biochemistry & Function

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The mechanism of placental alkaline phosphatase (PLAP) induction by prednisolone in a uterine cervical epidermoid cancer cell line SKG-IIIa was investigated in vitro by enzyme-cytochemistry, enzyme immunoassay, Northern and Southern blot analysis, and in situ hybridization. Enzyme-cytochemical alkaline phosphatase (ALP) staining and immunoassay revealed increased levels of PLAP (heat-stable ALP) in prednisolone-treated cells. Northern blot analysis and in situ hybridization showed increased amounts of PLAP mRNA. Southern blot analysis indicated that PLAP was not a product of an amplified or rearranged gene. These findings suggest that the induction of PLAP mRNA in SKG-IIIa cells by prednisolone in turn increased the levels of PLAP.

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Different lectin affinities in rat alkaline phosphatase isozymes: Multiple forms of the isozyme isolated by heterogeneities of sugar moieties

Cited by 39 publications

References 22 publications

Abnormal Glycosylation of Duodenal Membranous Alkaline Phosphatase Induced by Cysteamine in Rats

Abnormal Glycosylation of Duodenal Membranous Alkaline Phosphatase Induced by Cysteamine in Rats

Enhancement of drug delivery to bone: Characterization of human tissue-nonspecific alkaline phosphatase tagged with an acidic oligopeptide

The mechanism of placental alkaline phosphatase induction in vitro

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