ARTICLES

Jejunal phosphorylation and dephosphorylation of absorbed pyridoxine.HCl in vitro

The phosphorylation and dephosphorylation of absorbed vitamin was studied in rat jejunal everted sacs during in vitro uptake of [3H]pyridoxine.HCl. Kinetic studies during 4-min incubations demonstrated that phosphorylation was saturable (Km, 13.3 micron; Vmax, 0.906 nmol/4 min/g wet tissue) and was significantly inhibited by both structural and metabolic inhibitors. Longer incubations (30-min) demonstrated similar saturation and inhibition of net phosphorylation. Dephosphorylation of the phosphate esters formed by the phosphorylation step also occurred, as demonstrated by pulse-chase experiments. Disappearance of these phosphates was initially rapid, approximated first order kinetics, and apparently was mediated by a phosphatase. Whereas phosphorylation did not appear to affect initial uptake rates for [3H]pyridoxine.HCl by the jejunum, net phosphorylation after more prolonged incubations qualitatively paralleled accumulation of total absorbed vitamin in tissue over the same time. The primary effect of phosphorylation during jejunal uptake of pyridoxine by rat jejunum was to increase the accumulation of absorbed pyridoxine in tissue during prolonged incubations. Dephosphorylation had the net effect of limiting the magnitude of that accumulation.