Mammalian intestinal alkaline phosphatase acts as highly active exopolyphosphatase.
Recent results revealed that inorganic polyphosphates (polyP), being energy-rich linear polymers of orthophosphate residues known from bacteria and yeast, also exist in higher eukaryotes. However, the enzymatic basis of their metabolism especially in mammalian cells is still uncertain. Here we demonstrate for the first time that alkaline phosphatase from calf intestine (CIAP) is able to cleave polyP molecules up to a chain length of about 800. The enzyme acts as an exopolyphosphatase degrading polyP in a processive manner. The pH optimum is in the alkaline range. Divalent cations are not required for catalytic activity but inhibit the degradation of polyP. The rate of hydrolysis of short-chain polyP by CIAP is comparable to that of the standard alkaline phosphatase (AP) substrate p-nitrophenyl phosphate. The specific activity of the enzyme decreases with increasing chain length of the polymer both in the alkaline and in the neutral pH range. The K(m) of the enzyme also decreases with increasing chain length. The mammalian tissue non-specific isoform of AP was not able to hydrolyze polyP under the conditions applied while the placental-type AP and the bacterial (Escherichia coli) AP displayed polyP-degrading activity.
Lorenz
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B
Institute für Physiologische Chemie, Universität, Mainz, Germany. blorenz@mail.uni.mainz.de
Schröder
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HC
D017766:Acid Anhydride Hydrolases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000469:Alkaline Phosphatase / Q000378:metabolism*; D000818:Animals; D002413:Cations, Divalent; D002417:Cattle; D003035:Cobalt / Q000494:pharmacology; D004492:Edetic Acid / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007422:Intestines / Q000201:enzymology*; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology; D009532:Nickel / Q000494:pharmacology; D009596:Nitrophenols / Q000378:metabolism; D009943:Organophosphorus Compounds / Q000378:metabolism; D011122:Polyphosphates / Q000737:chemistry / Q000378:metabolism; D013379:Substrate Specificity
D002413:Cations, Divalent; D009596:Nitrophenols; D009943:Organophosphorus Compounds; D011122:Polyphosphates; C008644:nitrophenylphosphate; D003035:Cobalt; D009532:Nickel; D004492:Edetic Acid; D000469:Alkaline Phosphatase; D017766:Acid Anhydride Hydrolases; C020137:exopolyphosphatase; D008274:Magnesium