15-HETE: Selective incorporation into inositol phospholipids of MDCK cells

Abstract

The interaction of 15-hydroxyeicosatetraenoic acid (15-HETE) and cultured MDCK renal tubular epithelial cells was investigated to determine whether incorporation of this lipoxygenase product will affect polyphosphoinositide formation. MDCK cells were incubated with 1 microM [3H]-15-HETE for 15 to 120 minutes. Maximum uptake occurred between 15 and 30 minutes, and after 60 minutes, 70% of the incorporated 15-HETE was present in the phosphatidylinositol (PI) fraction. Some 15-HETE was also incorporated into phosphatidylinositol-4-monophosphate (PIP) and phosphatidylinositol-4,5-bisphosphate (PIP2). However, even though more 15-HETE than arachidonic acid was incorporated into PI, the fractional amount of 15-HETE present in the polyphosphoinositides was smaller than arachidonic acid. Therefore, although 15-HETE is selectively channeled into PI, conversion of PI species containing 15-HETE to PIP and PIP2 is relatively impaired. This suggests that either PI containing 15-HETE is a less effective substrate for phosphorylation, or PI containing arachidonic acid is a preferred substrate. MDCK cells converted 15-HETE to polar metabolites that were released into the extracellular fluid. This process may constitute a renal tubular mechanism for the clearance of 15-HETE and related lipoxygenase products.