The insulinomimetic effects of the polar head group of an insulin-sensitive glycophospholipid on pyruvate dehydrogenase in both subcellular and whole cell assays

Abstract

The polar head group that was released by treating an insulin-sensitive glycophospholipid with a phosphatidylinositol-specific phospholipase C (PI-PLC) stimulated pyruvate dehydrogenase (PDH) in both subcellular and whole cell assays. Stimulation of PDH activity in the subcellular assay was detected after gel filtration chromatography of the polar head group. This stimulation was not due to the presence of contaminating calcium and magnesium. The PDH-stimulating activity was proportional to the amount of polar head group added to the assay. The effect of the polar head group on PDH in the subcellular assay was blocked by sodium fluoride, suggesting that the polar head group activated the PDH phosphatase. In the whole cell assay, the polar head group stimulated PDH activity to an equal or greater extent as a physiological concentration of insulin. The effect of the polar head group was detected at 5 min, peaked at 10 min, and declined thereafter. In contrast, insulin stimulated PDH activity more slowly, but consistently. The PDH-stimulating activity eluted after bacitracin but ahead of ATP during gel filtration chromatography, and it was destroyed by exposure to NH 4OH or alkaline phosphatase and by boiling in water. These data support the proposal that an early step in insulin action is the release of insulinomimetic polar head group from the insulin-sensitive glycophospholipid.