The inhibition and disposition of intestinal alkaline phosphatase.
A primary mechanism of amino acid inhibition of intestinal alkaline phosphatase is postulated to be the formation of a dissociable enzyme-amino acid complex at an allosteric zinc site. The degree of inhibition was highly correlated with the Zn2+ stability constant of each amino acid and the inhibition was reversible by the addition of exogenous Zn2+ or by dialysis. This allosteric amino acid inhibition proved to be a useful probe of the membrane arrangement of the enzyme in the intact tissue. The catalytic site appears to face the lumen based on the poor permeability of the substrate, the accumulation of the coproducts in the luminal bath, and the response of the enzyme to luminal pH. Amino acid inhibition of alkaline phosphatase in the intact tissue was only effective in the presence of sodium; whereas sodium was not required in butanol extracted preparations which lacked the sidedness of the intact tissue. Since amino acid uptake from the intestine is sodium dependent, the allosteric inhibitory site is probably intracellular. The results suggest that the intestinal alkaline phosphatase spans the apical membrane with the catalytic site accessible from the lumen and the allosteric inhibitory site from the cytoplasm.
Gasser
K W
KW
Department of Zoology, Washington State University, Pullman 99164-4220.
Kirschner
L B
LB
D000469:Alkaline Phosphatase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000495:Allosteric Site; D000596:Amino Acids / Q000378:metabolism / Q000494:pharmacology*; D000818:Animals; D001665:Binding Sites; D003545:Cysteine / Q000378:metabolism / Q000494:pharmacology; D006639:Histidine / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007422:Intestines / Q000201:enzymology*; D007700:Kinetics; D010649:Phenylalanine / Q000378:metabolism / Q000494:pharmacology; D012487:Salmonidae / Q000378:metabolism*; D014337:Trout / Q000378:metabolism*; D015032:Zinc / Q000378:metabolism / Q000494:pharmacology
D000596:Amino Acids; D010649:Phenylalanine; D006639:Histidine; D000469:Alkaline Phosphatase; D015032:Zinc; D003545:Cysteine