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Expression, Solubilization, and Purification of Eukaryotic Borate Transporters

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Mouse GLUT9: evidences for a urate uniporter.

Stéphanie Bibert¹, Solange Kharoubi Hess, Dmitri Firsov

¹Dept. of Pharmacology and Toxicology, Univ. of Lausanne, Rue du Bugnon 27, CH-1005 Lausanne, Switzerland.

American Journal of Physiology. Renal Physiology

|July 10, 2009

Summary

Mouse GLUT9 (SLC2A9) functions as a high-capacity urate transporter, likely as an anion uniporter. Its splice variants show distinct kidney expression patterns, potentially explaining species differences in urate handling.

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Area of Science:

Molecular Biology
Physiology
Renal Physiology

Background:

GLUT9 (SLC2A9) is a recently identified urate transporter with incompletely understood functions.
Previous studies on human GLUT9 in Xenopus oocytes left its transport mechanism largely undefined.

Purpose of the Study:

To comprehensively characterize mouse GLUT9 transport properties, including its two splice variants (mGLUT9a and mGLUT9b).
To investigate the sensitivity of mouse GLUT9 to uricosuric drugs and determine its transport mechanism.
To analyze the kidney expression patterns of mGLUT9a and mGLUT9b.

Main Methods:

Utilized Xenopus laevis oocyte expression system for [(14)C]urate uptake assays.
Performed electrophysiological measurements to assess transport characteristics.
Employed RT-PCR on microdissected mouse kidney tubules to determine splice variant expression.

Main Results:

Both mGLUT9a and mGLUT9b demonstrated high-capacity urate transport with a K(m) of ~650 µM.
Uricosuric agents benzbromarone and losartan significantly inhibited urate uptake, as did phloretin.
Electrophysiology indicated electrogenic, voltage-dependent urate transport independent of Na(+) and Cl(-).
mGLUT9a was found in proximal tubules, while mGLUT9b was specifically expressed in distal convoluted and connecting tubules.

Conclusions:

Mouse GLUT9 likely functions as a urate (anion) uniporter.
The distinct kidney localization of mGLUT9a and mGLUT9b suggests specialized roles in urate reabsorption.
Observed species-specific expression differences in mouse versus human GLUT9 may explain variations in urate handling.