Membrane interactions of rat intestinal alkaline phosphatase: role of polar head groups.
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.
D000469:Alkaline Phosphatase / Q000378:metabolism*; D000818:Animals; D004134:Dimyristoylphosphatidylcholine; D007413:Intestinal Mucosa / Q000201:enzymology*; D008081:Liposomes; D008871:Microvilli / Q000201:enzymology*; D008968:Molecular Conformation; D010206:Papain; D010713:Phosphatidylcholines / Q000494:pharmacology*; D010739:Phospholipase D / Q000378:metabolism; D010743:Phospholipids / Q000494:pharmacology*; D051381:Rats; D013329:Structure-Activity Relationship; D010738:Type C Phospholipases / Q000378:metabolism
D008081:Liposomes; D010713:Phosphatidylcholines; D010743:Phospholipids; D000469:Alkaline Phosphatase; D010738:Type C Phospholipases; D010739:Phospholipase D; D010206:Papain; D004134:Dimyristoylphosphatidylcholine