Ecto-diadenosine 5',5'''-P1,P4-tetraphosphate (Ap4A)-hydrolase is expressed as an ectoenzyme in a variety of mammalian and human cells and adds new aspects to the turnover of Ap4A.
Ap4A and other dinucleotides participate in the regulation of hemostasis and blood pressure control. With the exception of two previously reported surface anchored ectoAp4A-hydrolases on bovine aortic endothelial and chromaffine cells, all Ap4A-hydrolases reported are intracellular or freely soluble. We demonstrated that ectoAp4A-hydrolases are present on a broad variety of cell types of different species: rat mesangial, bovine corneal epithelial, human Hep-G2 and peridontal cells. Ectoenzyme properties were evaluated on rat mesangium cells. Chromatography of purified plasma membranes on Sephacel 300 resulted in enrichment of ectoAp4A-hydrolase and in separation from ectoATPase. In contrast to ATPase, Ap4A-hydrolase was stable at room temperature. EctoAp4A-hydrolase also recognized ATP as substrate, and therefore is not highly specific. The molecular weight was 180 kD. Unlike ectoAMPase ectoAp4A-hydrolase was not attached via a glycosyl-phosphatidylinositol (GPI)-moiety. Concentrations of PI-PLC 10-100-fold higher than effective for ectoAMPase cleavage (10-100 mU/ml) plus extensively extended incubation times up to eight hours did not result in cleavage of ectoAp4A-hydrolase. The enzyme ectoAp4A-hydrolase might presage a direction for pharmaceutical manipulation in the control of blood pressure and hemostasis.
von Drygalski
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The Blood Center of Southeastern Wisconsin, Blood Research Institute, Milwaukee 53226-2178, USA. avdrygalski@bcsew.edu
Ogilvie
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D017766:Acid Anhydride Hydrolases / Q000378:metabolism*; D000818:Animals; D001011:Aorta; D002417:Cattle; D002478:Cells, Cultured; D019439:Chromaffin Cells / Q000201:enzymology / Q000378:metabolism*; D015226:Dinucleoside Phosphates / Q000378:metabolism*; D004730:Endothelium, Vascular / Q000201:enzymology / Q000378:metabolism*; D019573:Epithelium, Corneal / Q000201:enzymology / Q000378:metabolism*; D005920:Glomerular Mesangium / Q000201:enzymology / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D010519:Periodontium / Q000201:enzymology / Q000378:metabolism; D051381:Rats; D014407:Tumor Cells, Cultured
D015226:Dinucleoside Phosphates; C020733:diadenosine tetraphosphate; D017766:Acid Anhydride Hydrolases; C020023:bis(5'-nucleosyl)tetraphosphatase (asymmetrical)