Phosphoprotein phosphatase activity of bovine intestinal alkaline phosphatase

The phosphoprotein phosphatase activity of a commercial preparation of bovine intestinal alkaline phosphatase (EC 3.1.3.1) was examined using phosvitin and dentine phosphoprotein as substrates. Over 90% and 70% of the phosphorus from dentine phosphoprotein and phosvitin were hydrolyzed in 2 h. The optimum pH of the enzyme for the dephosphorylation of phosvitin and dentine phosphoprotein was nearly 6. No protein phosphatase activity was observed when the alkaline phosphatases from bovine liver and pulp were investigated.     

Dissociation-constants of metal-ion-complexes with alkaline phosphatase from pig kidney

Summary Using metal-ion buffers it was possible to remove Zn²⁺, Mg²⁺ and Mn²⁺ ions of pig kidney alkaline phosphatase reversibly. The dissociation constants obtained are K_EMg:4·10^–7 M, K_EMn:4·10^–8 M and K_EZn:8·10^–13 M (22°C, pH:9.6, :0.07).Acknowledgement: The authors thank Dr.H. U. Wolf for helpful suggestions and valuable discussion and MissH. Köth for technical assistance.     

Nature of inhibition of rat testicular alkaline phosphatase by isatin

Summary Isatin has been found to inhibit rat testicular alkaline phosphatase (EC 3.1.3.1). That the inhibition is non-competitive as well as non-allosteric is evident from a) the hyperbolic curve relating inhibition as a function of inhibitor concentration; b) the small change in enthalpy, free energy and entropy; c) the number of isatin molecules associating with 1 molecule of the enzyme (n=1.29); and, d) the decrease in the values of both K_m and V_max in the presence of isatin.     

Nature of inhibition of rat testicular alkaline phosphatase by isatin

Isatin has been found to inhibit rat testicular alkaline phosphatase (EC 3.1.3.1). That the inhibition is non-competitive as well as non-allosteric is evident from a) the hyperbolic curve relating inhibition as a function of inhibitor concentration; b) the small change in enthalpy, free energy and entropy; c) the number of isatin molecules associating with 1 molecule of the enzyme (n=1.29); and, d) the decrease in the values of both Km and Vmax in the presence of isatin.     

Possible role of intestinal alkaline phosphatase activity in thiamine transport

Summary Thiamine deficiency caused a marked decrease of intestinal alkaline phosphatase (al-Pase) activity, but had no effect on the Ca⁺⁺-ATPase activity and Ca⁺⁺-absorption in rats. The al-Pase activity was significantly decreased 1 h after oral administration of ethanol at 0.5 and 2.5 g/kg. In contrast, Mg⁺⁺-, Ca⁺⁺- and (Na⁺+K⁺)-ATPase activities did not change after the administration of ethanol. These findings show that the al-Pase activity, unlike the Ca⁺⁺-ATPase activity, is not related to Ca⁺⁺-absorption. A possible role of al-Pase activity in the active transport of thiamine in the intestine was discussed.     

Dissociation-constants of metat-ion-complexes with alkaline phosphatase from pig kidney.

Using metal-ion buffers it was possible to remove Zn2+, Mg2+ and Mn2+ ions of pig kidney alkaline phosphatase reversibly. The dissociation constants obtained are KEMg: 4 X 10(-7) M, KEMn: 4 X 10(-8) M and KEZn: 8 X 10(-13) M (22 degrees C, pH: 9.6, mu: 0.07).     

Partial purification and some characteristics of hamster molar alkaline phosphatase

Summary A rapid 3-step method is given to purify partially hamster molar alkaline phosphatase. Molecular weight was 50,200 and isoelectric point 3.7. The alkaline phosphatases in the mesenchymal and ectodermal parts of the tooth are probably identical.     

Activation and specificity of alkaline phosphatase of a mineralizing collagen-rich system

Summary Alkaline phosphatase from tibia tendon ofMeleagris gallopavo L. was highly purified. The enzyme activation by different ions was measured. Mg²⁺ showed a high activation with a broader spectrum of phosphomonoester hydrolization. The in vivo Mg²⁺ concentration was an optimum for in vitro activation.Dedicated to Prof. Dr. G. Pfefferkorn on the occasion of his 65^th birthday.We thank Deutsche Forschungsgemeinschaft for support.     

Activation and specificity of alkaline phosphatase of a mineralizing collagen-rich system

Alkaline phosphatase from tibia tendon of Meleagris gallopavo L. was highly purified. The enzyme activation by different ions was measured. Mg2+ showed a high activation with a broader spectrum of phosphomonoester hydrolization. The in vivo Mg2+ concentration was an optimum for in vitro activation.     

Intracellular localization and heterogeneity of alkaline phosphatase of Yoshida ascites sarcoma

Zusammenfassung In Yoshida-Sarkomzellen ist die alkalische Phosphatase vorwiegend in der partikelfreien Fraktion in gelöster Form lokalisiert. Gelfiltration fÜhrt zu einer Auftrennung des Enzyms im partikelfreien Überstand von 6 Fraktionen, des Enzyms in den Zellkernen bei 2 Fraktionen.

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We are grateful to Tenovus (Wales) for financial support.     

The indigogenic reaction for histochemical demonstration of alkaline and acid phosphatase

Résumé Le principe indigogénique à la détection histochimique des phosphates alcalins et acides a été appliqué. Ces substrates ont l'avantage de permettre une localisation précise de l'enzyme, sans ou avec minime diffusion. C'est aussi une méthode simple et directe pour la mise en évidence de ces enzymes sans passer par une réaction couplée. Un bisindigo très insoluble a lieu apparaît aux points d'activité enzymatique. Le 5-bromo-4-chlorocomposé donne un indigo bleu-vert tandis que le 5-bromo-6-chloro-composé donne un indigo magenta.     

Possible role of intestinal alkaline phosphatase activity in thiamine transport.

Thiamine deficiency caused a marked decrease of intestinal alkaline phosphatase (al-Pase) activity, but had no effect on the Ca++-ATPase activity and Ca++-absorption in rats. The al-Pase activity was significantly decreased 1 h after oral administration of ethanol at 0.5 and 2.5 g/kg. In contrast, Mg++-, Ca++-and (Na+ + K+)-ATPase activities did not change after the administration of ethanol. These findings show that the al-Pase activity, unlike the Ca++-ATPase activity, is not related to Ca++-absorption. A possible role of al-Pase activity in the active transport of thiamine in the intestine was discussed.     

An improved procedure for purification of alkaline phosphatase from rat kidney

Riassunto Viene descitto un nuovo metodo per la purificazione della fosfatasi alcalina da rene di ratto. La metodica comprende 4 tappe: (1) isolamento della frazione microsomiale; (2) solubilizzazione dell'enzima dalle strutture microsomiali per mezzo del desossicolato e della lipasi; (3) frazionamento cromatografico su DEAE-Sephadex A-25 ed eluzione con gradiente di concentrazione; (4) ricromatografia delle frazioni attive sullo stesso scambiatore anionico. All'elettroforesi su gel d'amido l'enzima ottenuto si comporta come una proteina omogenea.

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This work was supported by a grant from the Italian Consiglio Nazionale delle Ricerche, Impresa Enzimologia.