Characterization of structural and functional phosphoinositide domains in human erythrocyte membranes.
In the erythrocyte membrane, only a fraction (50-60%) of phosphatidylinositol 4,5-bisphosphate (PIP2) and of phosphatidylinositol 4-phosphate (PIP) is rapidly turned over by specific kinases and phosphatases and accessible to hydrolysis by the polyphosphoinositide (PPI)-specific phospholipase C (PLC). To investigate whether the metabolic segregation of PPI resulted from preferential interactions with proteins, we have measured the accessibility of PPI to bee venom phospholipase A2 (PLA2) in native erythrocyte membranes, or after treatments designed to remove peripheral proteins and cytoplasmic domains of integral proteins. In native membranes, PPI, as well as the other major phospholipids, behaved as two distinct fractions (R1 and R2) differing by their sensitivity to PLA2. Such a behavior was not observed in PIP and PIP2 containing artificial vesicles. Evidence was provided that the highly sensitive fraction of PIP and PIP2 (R1) may be identical to the PLC-sensitive and rapidly metabolized pool. Removal of peripheral proteins, followed by proteolysis of the cytoplasmic domain of integral proteins, mainly glycophorins and band 3, led to a reduction of the R1 fraction of PIP and of PIP2. It is proposed that the rapidly metabolized pool of PIP2 and PIP, involved in the regulation of major cellular functions, would be maintained in its functional state through interactions with integral proteins.
CNRS URA 1116, Université Paris XI, Orsay, France.
P
P
Gascard
M
M
Sauvage
J C
JC
Sulpice
F
F
Giraud
D001798:Blood Proteins / Q000737:chemistry; D004910:Erythrocyte Membrane / Q000737:chemistry* / Q000502:physiology; D006801:Humans; D008081:Liposomes; D008565:Membrane Proteins / Q000097:blood; D018129:Phosphatidylinositol Phosphates; D010716:Phosphatidylinositols / Q000097:blood* / Q000502:physiology; D010741:Phospholipases A; D054467:Phospholipases A2; D010738:Type C Phospholipases / Q000097:blood
D001798:Blood Proteins; D008081:Liposomes; D008565:Membrane Proteins; D018129:Phosphatidylinositol Phosphates; D010716:Phosphatidylinositols; D010741:Phospholipases A; D054467:Phospholipases A2; D010738:Type C Phospholipases