Functional characterization of mouse urea transporters UT-A2 and UT-A3 expressed in purified Xenopus laevis oocyte plasma membranes.
Urea is a small solute synthesized by many terrestrial organisms as part of the catabolism of protein. In mammals it is transported across cellular membranes by specific urea transporter (UT) proteins that are the products of two separate, but closely related genes, referred to as UT-A and UT-B. Three major UT-A isoforms are found in the kidney, namely UT-A1, UT-A2, and UT-A3. UT-A2 is found in the thin, descending limb of the loop of Henle, whereas UT-A1 and UT-A3 are concentrated in the inner medullary collecting duct. UT-A2 and UT-A3 effectively represent two halves of the whole UT-A gene and, when joined together by 73 hydrophilic amino acids, constitute UT-A1. A biophysical characterization of mouse UT-A2 and UT-A3 was undertaken by expression in Xenopus laevis oocytes and subsequent preparation of highly enriched plasma membrane vesicles for use in stopped-flow fluorometry. Both isoforms were found to be highly specific for urea, and did not permeate water, ammonia, or other molecules closely related to urea (formamide, acetamide, methylurea, and dimethylurea). Single transporter flux rates of 46,000 +/- 10,000 and 59,000 +/- 15,000 (means +/- SE) urea molecules/s/channel for UT-A2 and UT-A3, respectively, were obtained. Overall, the UT-A2 and UT-A3 isoforms appear to have identical functional kinetics.
Maciver
Bryce
B
Beth Israel Deaconess Medical Center and Harvard Medical School, 840 Memorial Drive, Cambridge MA 02139, USA. imaciver@bidmc.harvard.edu
Smith
Craig P
CP
Hill
Warren G
WG
Zeidel
Mark L
ML
D000818:Animals; D030801:Animals, Genetically Modified; D002462:Cell Membrane / Q000502:physiology*; D005260:Female; D008138:Loop of Henle / Q000502:physiology; D026901:Membrane Transport Proteins / Q000235:genetics* / Q000502:physiology; D051379:Mice; D009865:Oocytes / Q000502:physiology*; D014162:Transfection; D014982:Xenopus laevis; D000096982:Urea Transporters
D026901:Membrane Transport Proteins; D000096982:Urea Transporters