Phase partitioning detects differences between phospholipase-released forms of alkaline phosphatase--a GPI-linked protein.
A number of enzymes are known to release alkaline phosphatase and other glycan phosphatidylinositol-anchored proteins from membrane surfaces. We describe a novel approach to detect and measure these activities by partitioning in aqueous phase systems. The procedures avoid the complications of micelle-formation involving hydrophobic molecules that may arise with detergent-based partition systems and can clearly distinguish between inositol-specific phospholipase C and D activities.
Raymond
F D
FD
Department of Chemical Pathology, Royal Postgraduate Medical School, London, UK.
Moss
D W
DW
Fisher
D
D
D000469:Alkaline Phosphatase / Q000302:isolation & purification*; D001409:Bacillus cereus / Q000201:enzymology; D003911:Dextrans; D017261:Glycosylphosphatidylinositols / Q000737:chemistry*; D006801:Humans; D007527:Isoenzymes / Q000302:isolation & purification*; D043265:Phosphatidylinositol Diacylglycerol-Lyase; D010739:Phospholipase D / Q000737:chemistry; D010727:Phosphoric Diester Hydrolases / Q000302:isolation & purification; D010920:Placenta / Q000378:metabolism; D011092:Polyethylene Glycols; D012995:Solubility; D013302:Streptomyces / Q000201:enzymology; D010738:Type C Phospholipases / Q000737:chemistry*
D003911:Dextrans; D017261:Glycosylphosphatidylinositols; D007527:Isoenzymes; D011092:Polyethylene Glycols; D000469:Alkaline Phosphatase; D010727:Phosphoric Diester Hydrolases; D010738:Type C Phospholipases; D010739:Phospholipase D; D043265:Phosphatidylinositol Diacylglycerol-Lyase