Endogenous calcium in sickle cells does not activate polyphosphoinositide phospholipase C.
Sickle-cell-anaemia erythrocytes (SS cells) are known to have a high Ca2+ content (particularly the dense cell fraction) and to take up Ca2+ on deoxygenation. It has been reported that this high Ca2+ was responsible for the activation of the Ca2+-dependent K+ loss, and of the Ca2+-sensitive polyphosphoinositide phospholipase C (PIC) in dense SS cells. We found that, either in the total population of SS cells or in the light or dense fractions, the content of phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P2] was not changed, whereas that of phosphatidylinositol 4-phosphate was increased and that of phosphatidic acid (PtdOH) was decreased compared with normal (AA) erythrocytes. Deoxygenation-induced Ca2+ entry into SS cells did not change the concentration or, in 32P-prelabelled cells, the radioactivity of polyphosphoinositides and PtdOH. It also failed to induce the formation of inositol 1,4,5-trisphosphate, the product of PtdIns(4,5)P2 hydrolysis by PIC, which was measured by an original method using ion-pair reverse-phase h.p.l.c. Thus there was no evidence of an endogenous Ca2+ effect on the PIC activity in SS cells, in agreement with the demonstration that the excess Ca2+ in SS cells is compartmentalized into internal vesicles and unavailable as free Ca2+. The 32P incorporation in polyphosphoinositides and PtdOH was markedly higher in SS than in AA cells, but this increase was the same in both dense and light SS cells. The increase in the turnover of these phospholipids in SS cells is consistent either with an activation of the lipid kinases and phosphatases or with perturbation in the metabolic compartmentation of these lipids.
Rhoda
M D
MD
INSERM U 91, Hôpital Henri Mondor, Creteil, France.
Sulpice
J C
JC
Gascard
P
P
Galacteros
F
F
Giraud
F
F
D000255:Adenosine Triphosphate / Q000097:blood; D000755:Anemia, Sickle Cell / Q000097:blood / Q000201:enzymology*; D002118:Calcium / Q000097:blood*; D002851:Chromatography, High Pressure Liquid; D004912:Erythrocytes / Q000201:enzymology; D004913:Erythrocytes, Abnormal / Q000201:enzymology*; D006801:Humans; D015544:Inositol 1,4,5-Trisphosphate; D007295:Inositol Phosphates / Q000097:blood; D008055:Lipids / Q000097:blood; D010084:Oxidation-Reduction; D010712:Phosphatidic Acids / Q000097:blood; D010716:Phosphatidylinositols / Q000097:blood; D054801:Phosphoinositide Phospholipase C; D010743:Phospholipids / Q000097:blood; D010727:Phosphoric Diester Hydrolases / Q000097:blood*; D010761:Phosphorus Radioisotopes
D007295:Inositol Phosphates; D008055:Lipids; D010712:Phosphatidic Acids; D010716:Phosphatidylinositols; D010743:Phospholipids; D010761:Phosphorus Radioisotopes; D015544:Inositol 1,4,5-Trisphosphate; D000255:Adenosine Triphosphate; D010727:Phosphoric Diester Hydrolases; D054801:Phosphoinositide Phospholipase C; D002118:Calcium