ium-independent carrier-mediated inositol transport in cultured renal epithelial (LLC-PK_1) cells

摘要

In addition to the concentrative, Na+-dependent inositol transport system demonstrated in many cell types, carrier-mediated, Na+-independent inositol transport is also shown to exist in LLC-PK1 renal epithelia. Inhibition of inositol uptake in Na+-free saline by 0.1 mM phloretin, and self-inhibition by net concentrations of inositol exceeding 10 mM, demonstrate the carrier-mediation of the Na+-independent uptake and distinguish it from flux through anion channels. The Na+-dependent uptake exhibits higher affinity for inositol, as seen by the stronger self-inhibition at lower inositol concentrations in Na+ saline. Kinetic analyses indicate a Km of 178 μM and a Vmax of 2447 pmol/min per μg DNA for the Na+-dependent system, whereas the lower affinity, lower capacity Na+-independent system manifests a Km of 5.2 mM and a Vmax of 249 pmol/min per μg DNA. The Na+-independent uptake further differs from the Na+-dependent transport by the lack of inhibitory effect of 10 mM glucose, and the greater relative inhibition of phloretin compared to that of phlorizin. Both types of uptake appear to localize predominantly to the basal-lateral cell surface. The Na+-independent transport is bidirectional, functioning in efflux as well as influx of inositol.