Membrane interactions of rat intestinal alkaline phosphatase: role of polar head groups
- 1 November 1985
- journal article
- research article
- Published by American Chemical Society (ACS) in Biochemistry
- Vol. 24 (23), 6603-6608
- https://doi.org/10.1021/bi00344a045
Abstract
Lipid-protein interactions with purified membranous intestinal alkaline phosphatase have been studied by using rat intestine. The enzyme was incorporated equally well into neutral lecithin and anionic liposomes, including those made from phosphatidic acid alone. It could not be solubilized with chaotropic salts nor by phospholipases C and D from either native membranes or phospholipid vesicles. Detergents effected nearly complete release of enzyme from the vesicles. Phosphatase activity was lost upon treatment with phospholipase D alone. The activity was restored with free choline, or choline containing phospholipids, but not by the addition of other phospholipids or amines. The catalytic activity was also lower when the enzyme was bound to a phosphatidylcholine vesicle containing additional phosphatidic acid. Neither phosphatidylserine nor phosphatidylinositol addition altered enzyme activity. These results show that the enzyme binds to the membrane by a primary hydrophobic interaction with membrane phospholipids without requiring the polar head group and that the enzyme activity is affected via a secondary interaction with choline. We suggest that choline protects the active site of brush border alkaline phosphatase from inhibition by endogenous membrane phosphate groups.Keywords
This publication has 15 references indexed in Scilit:
- The mode of association of the enzyme complex sucrase.isomaltase with the intestinal brush border membrane.Journal of Biological Chemistry, 1979
- Phospholipases as probes for membrane sideness. Selective analysis of the outer monolayer of asymmetric bilayer vesiclesJournal of Biological Chemistry, 1978
- Integration of alkaline phosphatase in the intestinal brush border membraneBiochimica et Biophysica Acta (BBA) - Biomembranes, 1978
- A modified procedure for the rapid preparation of efficiently transporting vesicles from small intestinal brush border membranes. Their use in investigating some properties of d-glucose and choline transport systemsBiochimica et Biophysica Acta (BBA) - Biomembranes, 1978
- Release of alkaline phosphatase from membranes by a phosphatidylinositol-specific phospholipase C.Biochemical Journal, 1977
- Modification of erythrocyte membranes by a purified phosphatidylinositol-specific phospholipase C (Staphylococcus aureus)Biochemical Journal, 1977
- Phosphorus Assay in Column ChromatographyJournal of Biological Chemistry, 1959
- A SIMPLE METHOD FOR THE ISOLATION AND PURIFICATION OF TOTAL LIPIDES FROM ANIMAL TISSUESJournal of Biological Chemistry, 1957
- Mechanism of action of alkaline phosphataseArchives of Biochemistry and Biophysics, 1952
- PROTEIN MEASUREMENT WITH THE FOLIN PHENOL REAGENTJournal of Biological Chemistry, 1951