Research Articles
Modulation of sulfate renal transport by alterations in cell membrane fluidity

https://doi.org/10.1021/js990114cGet rights and content

Abstract

Changes in membrane fluidity have been shown to alter the sodium‐dependent renal transport of glucose and phosphate; however, this has not been examined for sodium/sulfate cotransport in the renal proximal tubule. Sodium/sulfate cotransport regulates the homeostasis of sulfate in mammals. The objective of this study was to investigate the influence of alterations of membrane fluidity on sodium‐coupled sulfate transport in the Madin–Darby canine kidney cells, which have been stably transfected with sodium/sulfate cotrans‐porter (NaSi‐1) cDNA (MDCK‐Si). Preincubation of cells with 0.2 mM cholesterol significantly decreased the Vmax for sodium/sulfate cotrans‐port (13.69 ±1.11 vs 10.15 ±1.17 nmol/mg protein/5 min, mean ±SD, n = 4, p < 0.01) with no significant alteration in Km. The addition of benzyl alcohol (20 mM) to cells increased the Vmax of sulfate uptake by 20% (11.97 ±0.91 vs 14.35 ±0.56 nmol/mg protein/5 min, mean ±SD, n = 3, p < 0.05) with no significant change in Km. Membrane fluidity, as measured by the fluorescence polarization of 1,6‐diphenyl 1,3,5‐hexatriene (DPH), was significantly increased in MDCK‐Si cells treated with 20 mM benzyl alcohol and decreased in the cells preincubated with 0.2 mM cholesterol, compared with control cells. Our results suggest that alterations in membrane fluidity that may occur as a result of disease states, aging, and pregnancy may play an important role in the modulation of renal sodium/sulfate cotransport.

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