title: A physiologic function for alkaline phosphatase: endotoxin detoxification.
abstract: Alkaline phosphatase (AP), a common enzyme present in many species including humans, has been studied extensively. Although the enzyme is routinely applied as a marker for liver function, its biologic relevance is poorly understood. The reason for this is obvious: the pH optimum of AP in vitro, as measured with the usual test substrates (+/-10.5), greatly exceeds the physiologic pH range as it occurs in biologic tissues. We now hypothesize that this relatively high pH optimum in vitro is related to dissociation of acidic groups in the protein preparation, which leads to the formation of negatively charged groups in the vicinity of the active site of the enzyme. These negatively charged groups may promote the activity of AP. We examined the possibility that endotoxin is a natural substrate for this enzyme because this phosphorylated substance is able to supply multiple negatively charged residues in the microenvironment of the enzyme at a physiologic pH level. Phosphate groups in the endotoxin molecule are known to be essential for the biologic activities of this bacterial product. The present study demonstrates that in intestinal and renal tissue specimens in vitro, AP is endowed with endotoxin dephosphorylating activity at pH levels closer to the physiologic range. This is also illustrated by our experiments in vivo showing that the toxicity of endotoxin is significantly reduced after exposure to AP preparations, as tested by inducing a local intradermal inflammatory reaction in rats. Collectively, our data suggest that the ubiquitous enzyme AP may accomplish protection against endotoxin, an equally ubiquitous product of Gram-negative bacteria that may cause lethal complications after an infection with these micro organisms.
authors.affiliation: Department of Pharmacokinetics and Drug Delivery, University of Groningen, The Netherlands.
authors.forename: K
authors.initials: K
authors.lastname: Poelstra
authors.forename: W W
authors.initials: WW
authors.lastname: Bakker
authors.forename: P A
authors.initials: PA
authors.lastname: Klok
authors.forename: M J
authors.initials: MJ
authors.lastname: Hardonk
authors.forename: D K
authors.initials: DK
authors.lastname: Meijer
mesh_terms: D000469:Alkaline Phosphatase / Q000378:metabolism*; D000818:Animals; D004731:Endotoxins / Q000493:pharmacokinetics* / Q000633:toxicity; D004926:Escherichia coli; D006863:Hydrogen-Ion Concentration; D008658:Inactivation, Metabolic; D007413:Intestinal Mucosa / Q000201:enzymology*; D007668:Kidney / Q000201:enzymology*; D007672:Kidney Cortex / Q000201:enzymology; D007684:Kidney Tubules / Q000201:enzymology; D007700:Kinetics; D008297:Male; D008871:Microvilli / Q000201:enzymology; D051381:Rats; D011919:Rats, Inbred Strains
chemical_list: D004731:Endotoxins; D000469:Alkaline Phosphatase