The alkaline phosphatase from bone: transphosphorylating activity and kinetic mechanism

Müller, Karin; Schellenberger, Volker; Borneleit, Petra; Treide, A

doi:10.1016/0167-4838(91)90283-6

Cited by 22 publications

(9 citation statements)

References 19 publications

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“…The data are in agreement with the inhibition by levamisole at alkaline pH of purified L/B/K ALP [Van Belle, 1976] and L/B/K ALP measured in extracts of P19 and HL-60 cells [Scheibe et al, 1991;Wei et al, 1992] (R.J. Scheibe, unpublished observation). At pH 7.4, levamisole also inhibited activity of purified or membrane-bound ALP of chick cartilage, bone, and rat liver [Cyboron et al, 1982;Farley et al, 1982;Mü ller et al, 1991;Sarrouilhe et al, 1993]. Levamisole did not inhibit extracellular phosphohydrolase activities of other membrane enzymes [El Kouni and Cha, 1982;LeBel et al, 1980], pointing out the specificity of levamisole toward ALP.…”

Section: Discussionmentioning

confidence: 99%

Ecto-alkaline phosphatase activity identified at physiological pH range on intact P19 and HL-60 cells is induced by retinoic acid

et al. 2000

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The activity of membrane-bound alkaline phosphatase (ALP) expressed on the external surface of cultured murine P19 teratocarcinoma and human HL-60 myeloblastic leukemia cells was studied at physiological pH using p-nitrophenylphosphate (pNPP) as substrate. The rate of substrate hydrolysis catalyzed by intact viable cells remained constant for eight successive incubations of 30 min and was optimal at micromolar substrate concentrations over the pH range 7.4-8.5. The value of apparent K(m) for pNPP in P19 and HL-60 cells was 120 microM. Hydrolytic activity of the ecto-enzyme at physiological pH decreased by the addition of levamisole, a specific and noncompetitive inhibitor of ALP (K(i) P19 = 57 microM; K(i) HL-60 = 50 microM). Inhibition of hydrolysis was reversed by removal of levamisole within 30 min. Retinoic acid (RA), which promotes the differentiation of P19 and HL-60 cells, induced levamisole-sensitive ecto-phosphohydrolase activity at pH 7.4. After its autophosphorylation by ecto-kinase activity, a 98-kDa membrane protein in P19 cells was found to be sensitive to ecto-ALP, and protein dephosphorylation increased after incubation of cells with RA for 24 h and 48 h. Orthovanadate, an inhibitor of all phosphatase activities, blocked the levamisole-sensitive dephosphorylation of the membrane phosphoproteins, while (R)-(-)-epinephrine reversed the effect by complexation of the inhibitor. The results demonstrate that the levamisole-sensitive phosphohydrolase activity on the cell surface is consistent with ecto-ALP activity degrading both physiological concentrations of exogenously added substrate and endogenous surface phosphoproteins under physiological pH conditions. The dephosphorylating properties of ecto-ALP are induced by RA, suggesting a specific function in differentiating P19 teratocarcinoma and HL-60 myeloblastic leukemia cells.

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Section: Discussionmentioning

confidence: 99%

Ecto-alkaline phosphatase activity identified at physiological pH range on intact P19 and HL-60 cells is induced by retinoic acid

et al. 2000

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“…Conservation of alkaline phosphatase expression hints at an important role, but to date there is little biochemical evidence for alkaline phosphatase function except in bone mineralization (Yoon et al 1989;Beertsen and van den Bos 1992;Fedde et al 1999) and dephosphorylation of adenosine nucleotides (Gallo et al 1997). The alkaline phosphatases have been postulated to be involved in a range of other processes, including cell adhesion (Hui et al 1993), vitamin B transport (Rindi et al 1995), metastasis (Manara et al 2000), and cell signaling (Swarup et al 1981;Muller et al 1991). We are interested in their role in the development of mammalian preimplantation embryos.…”

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confidence: 99%

Inhibitor Profiles of Alkaline Phosphatases in Bovine Preattachment Embryos and Adult Tissues

McDougall

Plumb

King

et al. 2002

J Histochem Cytochem.

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The alkaline phosphatases are a small family of isozymes. Bovine preattachment embryos transcribe mRNA for two tissue-specific alkaline phosphatases (TSAP2 and TSAP3) beginning at the 4- and 8-cell stages. Whereas no mRNA has been detected in oocytes, there is maternally inherited alkaline phosphatase activity. It is not known which isozyme(s) is responsible for the maternal activity or when TSAP2 and TSAP3 form functional protein. No antibodies are available that recognize the relevant bovine alkaline phosphatases. Therefore, sensitivity to heat and chemical inhibition was used to separate the different isozymes. By screening tissues, it was determined that the bovine tissue-nonspecific alkaline phosphatase (TNAP) is inactivated by low temperatures (65C) and low concentrations of levamisole (<1 mM), whereas bovine tissue-specific isozymes require higher temperatures (90C) and levamisole concentrations (>5 mM). Inhibition by L-homoarginine and L-phenylalanine was less informative. Cumulus cells transcribe two isozymes and the pattern of inhibition suggested heterodimer formation. Inhibition of alkaline phosphatase in bovine embryos before the 8-cell stage indicated the presence of only TNAP. At the 16-cell stage the pattern was consistent with TNAP plus TSAP2 or -3 activity, and in morulae and blastocysts the pattern indicated that the maternal TNAP is fully supplanted by TSAP2 or TSAP3.

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“…It involves the activation of a serine by a zinc atom, the formation of a phosphorylenzyme, the hydrolysis of the phosphoseryl by a water molecule activated by a second zinc atom and the release of the phosphate or its transfer to an acceptor. Four main catalytic functions have been attributed to these enzymes, hydrolase activity on low molecular weight phosphomonoesters [ 8 ], phosphotransferase activity [ 9 ], protein phosphatase activity [ 10 ] and pyrophosphatase activity [ 11 ]. The physiological role of AP is poorly known, except for the involvement of the NSAP isoenzyme in the mineralisation of bone tissue [ 12 ].…”

Section: Introductionmentioning

confidence: 99%

Kinetic comparison of tissue non-specific and placental human alkaline phosphatases expressed in baculovirus infected cells: application to screening for Down's syndrome

Denier

Biasini

et al. 2002

BMC Biochem

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Background In humans, there are four alkaline phosphatases, and each form exibits a characteristic pattern of tissue distribution. The availability of an easy method to reveal their activity has resulted in large amount of data reporting correlations between variations in activity and illnesses. For example, alkaline phosphatase from neutrophils of mothers pregnent with a trisomy 21 fetus (Down's syndrome) displays significant differences both in its biochemical and immunological properties, and in its affinity for some specific inhibitors. Results To analyse these differences, the biochemical characteristics of two isozymes (non specific and placental alkaline phosphatases) were expressed in baculovirus infected cells. Comparative analysis of the two proteins allowed us to estimate the kinetic constants of denaturation and sensitivity to two inhibitors (L-p-bromotetramisole and thiophosphate), allowing better discrimination between the two enzymes. These parameters were then used to estimate the ratio of the two isoenzymes in neutrophils of pregnant mothers with or without a trisomy 21 fetus. It appeared that the placental isozyme represented 13% of the total activity of neutrophils of non pregnant women. This proportion did not significantly increase with normal pregnancy. By contrast, in pregnancies with trisomy 21 fetus, the proportion reached 60–80% of activity. Conclusion Over-expression of the placental isozyme compared with the tissue-nonspecific form in neutrophils of mother with a trisomy 21 fetus may explain why the characteristics of the alkaline phosphatase in these cells is different from normal. Application of this knowledge could improve the potential of using alkaline phosphatase measurements to screen for Down's syndrome.

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The alkaline phosphatase from bone: transphosphorylating activity and kinetic mechanism

Cited by 22 publications

References 19 publications

Ecto-alkaline phosphatase activity identified at physiological pH range on intact P19 and HL-60 cells is induced by retinoic acid

Ecto-alkaline phosphatase activity identified at physiological pH range on intact P19 and HL-60 cells is induced by retinoic acid

Inhibitor Profiles of Alkaline Phosphatases in Bovine Preattachment Embryos and Adult Tissues

Kinetic comparison of tissue non-specific and placental human alkaline phosphatases expressed in baculovirus infected cells: application to screening for Down's syndrome

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