[Methodological study on the assay of glycosylphosphatidylinositol-specific phospholipase D activity in serum].
A method for the assay of glycosylphosphatidylinositol-specific phospholipase D(GPI-PLD) activity in human serum was established by using glycosylphosphatidylinositol(GPI) anchored placental alkaline phosphatase(PLAP) as a substrate. Serum GPI-PLD activity was determined both qualitatively and quantitatively by PEG/dextran and triton-X-114 partitioning respectively, and some kinetic properties of its enzymatic reaction were also studied. This method revealed a satisfactory reproductivity(CV = 3.6%) and a high sensitivity(2 microliters serum was sufficient). In addition, it is practical, economical, and feasible to be widely applied. The activities of GPI-PLD measured by this method in 100 normal subjects ranged from 30% to 50% (in terms of the percentage of converting GPI anchored PLAP substrate to the anchor-free product).
Tang
J
J
Department of Biochemistry, Hunan Medical University, Changsha 410078.
Li
W
W
D000328:Adult; D000469:Alkaline Phosphatase; D001681:Biological Assay / Q000379:methods; D006801:Humans; D008875:Middle Aged; D010739:Phospholipase D / Q000097:blood*; D010920:Placenta; D012016:Reference Values
D000469:Alkaline Phosphatase; D010739:Phospholipase D; C065017:glycoprotein phospholipase D