Urea transporter UT3 functions as an efficient water channel - Direct evidence for a common water/urea pathway

A family of molecular urea transporters (UTs) has been identified whose members appear to have an exceptionally high transport turnover rate. To test the hypothesis that urea transport involves passage through an aqueous channel, osmotic water permeability was measured in Xenopus oocytes expressing UTs, The UT3 class of urea transporters functioned as efficient water channels. Quantitative measurement of single channel water permeability (p(f)) using epitope-tagged rat UTs gave p(f) (in cm(3)/s x 10(-14)) of 0.14 +/- 0.11 (UT2) and 1.4 +/- 0.2 (UT3), compared with 6.0 and 2.3 for water channels AQP1 and AQP3, respectively. Relative single channel urea permeabilities (p(urea)) were 1.0 (UT2), 0.44 (UT3), and 0.0 (AQP1), UT3-mediated water and urea transport were weakly temperature-dependent (activation energy <4 kcal/mol), inhibited > 75% by the urea transport inhibitor 1,3-dimethylthiourea, but not inhibited by the water transport inhibitor HgCl2. To test for a common water/urea pore, the urea reflection coefficient (sigma(urea)) was measured by independent induced osmosis and solvent drag methods. In UT3-expressing oocytes, the time course of oocyte volume in response to different urea gradients (induced osmosis) gave sigma(urea) similar to 0.3 for the UT3 pathway, in agreement with sigma(urea) determined by the increase in uptake of [C-14]urea during osmotic gradient-induced oocyte swelling (solvent drag). In oocytes of comparable water and urea permeability coexpressing AQP1 (permeable to water, not urea) and UT2 (permeable to urea, not water), sigma(urea) = 1. These results indicate that UT3 functions as a urea/water channel utilizing a common aqueous pathway. The water transporting function and low urea reflection coefficient of UT3 in vasa recta may be important for the formation of a concentrated urine by countercurrent exchange in the kidney.