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Hoffmann, R. A wiki for the life sciences where authorship matters. Nature Genetics (2008)

Functional properties of the apical Na+-K+-2Cl- cotransporter isoforms.

The bumetanide-sensitive Na(+):K(+):2Cl(-) cotransporter (BSC1) is the major pathway for salt reabsorption in the apical membrane of the mammalian thick ascending limb of Henle. Three isoforms of the cotransporter, known as A, B, and F, exhibit axial expression along the thick ascending limb. We report here a functional comparison of the three isoforms from mouse kidney. When expressed in Xenopus oocytes the mBSC1-A isoform showed higher capacity of transport, with no difference in the amount of surface expression. Kinetic characterization revealed divergent affinities for the three cotransported ions. The observed EC(50) values for Na(+), K(+), and Cl(-) were 5.0 +/- 3.9, 0.96 +/- 0.16, and 22.2 +/- 4.8 mm for mBSC1-A; 3.0 +/- 0.6, 0.76 +/- 0.07, and 11.6 +/- 0.7 mm for mBSC1-B; and 20.6 +/- 7.2, 1.54 +/- 0.16, and 29.2 +/- 2.1 mm for mBSC1-F, respectively. Bumetanide sensitivity was higher in mBSC1-B compared with the mBSC1-A and mBSC1-F isoforms. All three transporters were partially inhibited by hypotonicity but to different extents. The cell swelling-induced inhibition profile was mBSC1-F > mBSC1-B > mBSC1-A. The function of the Na(+):K(+):2Cl(-) cotransporter was not affected by extracellular pH or by the addition of metolazone, 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS), or R(+)-[(2-n-butyl-6,7-dichloro-2-cyclopentyl-2,3-dihydro-1-oxo-1-H-indenyl-5-yl)-oxy]acetic acid (DIOA) to the extracellular medium. In contrast, exposure of oocytes to HgCl(2) before the uptake period reduced the activity of the cotransporter. The effect of HgCl(2) was dose-dependent, and mBSC1-A and mBSC1-B exhibited higher affinity than mBSC1-F. Overall, the functional comparison of the murine apical renal-specific Na(+):K(+):2Cl(-) cotransporter isoforms A, B, and F reveals important functional, pharmacological, and kinetic differences, with both physiological and structural implications.[1]


  1. Functional properties of the apical Na+-K+-2Cl- cotransporter isoforms. Plata, C., Meade, P., Vazquez, N., Hebert, S.C., Gamba, G. J. Biol. Chem. (2002) [Pubmed]
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