The inositol lipids of Paramecium tetraurelia and preliminary characterizations of phosphoinositide kinase activity in the ciliary membrane.
Inositol glycerolipids make up less than 10% of total phospholipids of Paramecium tetraurelia cells. Unlike inositol lipids found in mammalian and other cell types, these lipids from Paramecium lack arachidonic acid. It was demonstrated that kinase and possibly phosphatase enzymes that interconvert phosphatidylinositol (PI), phosphatidylinositol phosphate (PI-P) and phosphatidylinositol-bis-phosphate (PI-P2) exist in ciliary membranes of this ciliate. When exogenous soybean PI and [gamma-32P]ATP were provided as substrates, isolated cilia preparations exhibited PI and PI-P kinase activities as demonstrated by the incorporation of radiolabel into PI-P and PI-P2. Kinase activity was activated by millimolar [Mg2+] and inhibited by millimolar [Ca2+]. Significant inhibition of kinase activity in the presence of unlabeled excess ATP suggested that ATP is the preferred phosphate donor for this reaction. Of 4 suborganellar fractions of isolated cilia, the membrane fraction had the greatest kinase activity indicating that the enzyme(s) is membrane-associated.
Suchard
S J
SJ
Department of Biological Sciences, University of Cincinnati, Ohio 45221-0006.
Rhoads
D E
DE
Kaneshiro
E S
ES
D019870:1-Phosphatidylinositol 4-Kinase; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002855:Chromatography, Thin Layer; D002923:Cilia / Q000032:analysis / Q000201:enzymology*; D003720:Densitometry; D006160:Guanosine Triphosphate / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D010247:Paramecium / Q000032:analysis* / Q000201:enzymology / Q000648:ultrastructure; D010716:Phosphatidylinositols / Q000032:analysis*; D010766:Phosphorylation; D010770:Phosphotransferases / Q000378:metabolism*
D010716:Phosphatidylinositols; D006160:Guanosine Triphosphate; D000255:Adenosine Triphosphate; D010770:Phosphotransferases; D019870:1-Phosphatidylinositol 4-Kinase