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J. Biol. Chem., Vol. 264, Issue 12, 6817-6822, 04, 1989
TA Rooney, R Hager, E Rubin and AP Thomas
The ability of alcohols to regulate inositol lipid-specific phospholipase C
(phosphoinositidase C) was examined in turkey erythrocyte ghosts prepared
by cell lysis of erythrocytes which were prelabeled with [3H] inositol.
Guanosine 5'-[gamma-thiotriphosphate] GTP[S] stimulated the production of
both [3H]inositol bisphosphate (18- fold) and [3H]inositol trisphosphate
(6-fold) in this system. The accumulation of [3H]inositol bisphosphate and
[3H]inositol trisphosphate was linear up to 8 min following an initial lag
period of 1-2 min. Ethanol (300 mM) reduced the lag period for [3H]inositol
phosphate accumulation at submaximal GTP[S] concentrations and caused a
shift to the left (3-fold) in the dose-response curve. Other short chain
alcohols, methanol (300 mM), 1-propanol (200 mM), and 1-butanol (50 mM)
also enhanced the accumulation of [3H] inositol phosphates in the presence
of submaximal GTP[S] concentrations. Receptor activation by the purinergic
agonist adenosine 5'-[beta-thio]disphosphate (ADP[S]) (10 microM) also
reduced the lag period for [3H] inositol phosphate formation and shifted
the GTP[S] dose response to the left (10-fold). In addition, ADP[S]
increased the response to maximal GTP[S] concentrations. The formation of
[3H]inositol phosphates induced by GTP[S] was associated with a concomitant
decrease in labeling of both [3H]phosphatidylinositol monophosphate and
[3H]phosphatidylinositol bisphosphate, but no decrease in
[3H]phosphatidylinositol was observed. All of the alcohols tested enhanced
the breakdown of [3H]polyphosphoinositides in the presence of GTP[S]. The
dose response to guanosine 5'-[beta gamma-imino]triphosphate for
[3H]inositol phosphate formation was displaced to the left by ethanol (300
mM) and ADP[S] (10 microM) (2- and 7-fold), respectively. ADP[S] also
enhanced the maximal response to guanosine 5'-[beta
gamma-imino]triphosphate. The [3H]inositol phosphate formation produced in
response to NaF was unaffected by either ethanol or receptor activation.
These results indicate that alcohols initiate an activation of
phosphoinositidase C, mediated at the level of the regulatory guanine
nucleotide-binding protein.
Short chain alcohols activate guanine nucleotide-dependent phosphoinositidase C in turkey erythrocyte membranes
Department of Pathology and Cell Biology, Thomas Jefferson University, Philadelphia, Pennsylvania 19107.
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