Browsing by Author "Molloy, C J"
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Item The Calcium Signal for Balb/MK Keratinocyte Terminal Differentiation Induces Sustained Alterations in Phosphoinositide Metabolism without Detectable Protein Kinase C Activation.(Journal of Biological Chemistry, 1989) Moscat, Jorge; Fleming, T P; Molloy, C J; López Barahona, Mónica; Aaronson, S ABalb/MK keratinocytes require epidermal growth factor for proliferation and terminally differentiate in response to elevated extracellular Ca2+ concentrations. The molecular pathways controlling cell differentiation in this system have yet to be established. We show that a dramatic and sustained activation of phosphoinositide metabolism is produced upon addition of Ca2+ to Balb/MK cultures. The pattern of inositol trisphosphate isomers released in response to Ca2+ challenge appeared to be atypical. Inositol 1,3,4-trisphosphate release was observed by 30 s and was produced earlier than any alteration in inositol 1,4,5-trisphosphate levels. Concomitant with the liberation of inositol phosphates, an increased production of diacylglycerol was observed. Despite a 3-fold increase in diacylglycerol levels detected even at 12 h after Ca2+ addition, no evidence of functional activation or down-regulation of protein kinase C was found. This was established by measuring p80 phosphorylation, epidermal growth factor binding, and protein kinase C levels by immunoblotting. Analysis of the diacylglycerol generated following Ca2+ addition to Balb/MK cells revealed that a significant proportion of that lipid was an alkyl ether glyceride molecular species. Therefore, it is possible that this diacylglycerol molecular species may play a role in the Ca2+-induced differentiation program of Balb/MK cells through mechanisms other than stimulation of classical protein kinase C.