Solubilization of membrane-bound matrix-induced alkaline phosphatase with polyoxyethylene 9-lauryl ether (polidocanol): purification and metalloenzyme properties.
1. Matrix-induced alkaline phosphatase prepared from rat osseous plate was solubilized with polidocanol and purified on a Sephacryl S-300 column. 2. Purified solubilized alkaline phosphatase has a molecular weight of ca 115,000 and bind one magnesium and two zinc ions. At least 110 detergent molecules are bound to each enzyme molecule. 3. Solubilization and purification procedures did not destroy the ability of the enzyme to hydrolyze adenosine-5'-triphosphate, p-nitrophenylphosphate, pyrophosphate and bis p-nitrophenylphosphate. 4. Magnesium, manganese and cobalt ions are stimulators of PNPPase activity of solubilized enzyme whereas calcium and zinc ions are inhibitors.
Ciancaglini
P
P
Departamento de Química, Faculdade de Filosofia, Ribeirão Preto, Brasil.
Pizauro
J M
JM
Rezende
A A
AA
Rezende
L A
LA
Leone
F A
FA
D009597:4-Nitrophenylphosphatase / Q000378:metabolism; D000469:Alkaline Phosphatase / Q000302:isolation & purification* / Q000378:metabolism; D000818:Animals; D003902:Detergents; D006132:Growth Plate / Q000201:enzymology; D066298:In Vitro Techniques; D007700:Kinetics; D008566:Membranes / Q000201:enzymology; D008670:Metals / Q000494:pharmacology; D008970:Molecular Weight; D000077423:Polidocanol; D011092:Polyethylene Glycols; D051381:Rats; D012995:Solubility
D003902:Detergents; D008670:Metals; D000077423:Polidocanol; D011092:Polyethylene Glycols; D000469:Alkaline Phosphatase; D009597:4-Nitrophenylphosphatase