Characterization of phosphoinositide kinases in normal and sickle anaemia red cells.
PtdIns and PtdInsP kinases from normal erythrocyte (AA) membranes and sickle cell anaemia erythrocyte (SS) membranes have been characterized. PtdIns kinase was studied in native membranes under conditions in which PtdInsP kinase and PtdInsP phosphatase do not express any activity. Kinetic analysis of the AA and SS PtdIns kinases indicate similar Km values for PtdIns and ATP but higher Vmax values for SS PtdIns kinase. PtdInsP kinase was partially purified from erythrocyte ghosts by NaCl extraction. The kinetic parameters of PtdInsP kinase determined under these conditions were similar in AA and SS NaCl extracts. These data suggest the presence of some effector of PtdIns kinase in SS cell membranes, resulting in a greater activity of the enzyme. This leads consequently, to increase the PtdIns4P pool and to activate PtdInsP kinase, in agreement with our previous observations of a greater [32P]Pi incorporation in both polyphosphoinositides in SS cells relatively to AA cells.
INSERM U. 359 CHRU, Pointe-à-Pitre, Guadeloupe.
M D
MD
Rhoda-Hardy-Dessources
R S
RS
de Neef
G
G
Mérault
F
F
Giraud
D019870:1-Phosphatidylinositol 4-Kinase; D000241:Adenosine / Q000494:pharmacology; D000255:Adenosine Triphosphate / Q000378:metabolism; D000755:Anemia, Sickle Cell / Q000201:enzymology*; D002478:Cells, Cultured; D004910:Erythrocyte Membrane / Q000201:enzymology*; D004913:Erythrocytes, Abnormal / Q000201:enzymology*; D006493:Heparin / Q000494:pharmacology; D006801:Humans; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology; D010716:Phosphatidylinositols / Q000378:metabolism; D010770:Phosphotransferases / Q000378:metabolism*; D017853:Phosphotransferases (Alcohol Group Acceptor); D013096:Spermine / Q000494:pharmacology
D010716:Phosphatidylinositols; D013096:Spermine; D000255:Adenosine Triphosphate; D006493:Heparin; D010770:Phosphotransferases; D017853:Phosphotransferases (Alcohol Group Acceptor); D019870:1-Phosphatidylinositol 4-Kinase; C020077:1-phosphatidylinositol-4-phosphate 5-kinase; D008274:Magnesium; D000241:Adenosine