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3-Hydroxyglutaric acid is transported via the sodium-dependent dicarboxylate transporter NaDC3

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Abstract

Patients with glutaryl-CoA dehydrogenase (GCDH) deficiency accumulate glutaric acid (GA) and 3-hydroxyglutaric acid (3OH-GA) in their blood and urine. To identify the transporter mediating the translocation of 3OH-GA through membranes, kidney tissue of Gcdh−/− mice have been investigated because of its central role in urinary excretion of this metabolite. Using microarray analyses of kidney-expressed genes in Gcdh−/− mice, several differentially expressed genes encoding transporter proteins were identified. Real-time polymerase chain reaction analysis confirmed the upregulation of the sodium-dependent dicarboxylate cotransporter 3 (NaDC3) and the organic cation transporter 2 (OCT2). Expression analysis of NaDC3 in Xenopus laevis oocytes by the two-electrode-voltage-clamp technique demonstrated the sodium-dependent translocation of 3OH-GA with a K M value of 0.95 mM. Furthermore, tracer flux measurements in Chinese hamster ovary cells overexpressing OCT2 showed that 3OH-GA inhibited significantly the uptake of methyl-4-phenylpyridinium, whereas 3OH-GA is not transported by OCT2. The data demonstrate for the first time the membrane translocation of 3OH-GA mediated by NaDC3 and the cis-inhibitory effect on OCT2-mediated transport of cations.

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Abbreviations

GA1:

glutaric aciduria type 1

GA:

glutaric acid

Gcdh:

glutaryl-CoA dehydrogenase

NaDC3:

sodium-dependent dicarboxylate cotransporter 3

3OH-GA:

3-hydroxyglutaric acid

OCT2:

organic cation transporter 2

Slc:

solute carrier

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Acknowledgements

This work was supported by the Deutsche Forschungsgemeinschaft (GRK 336 to Keyser and Stellmer; grant BU998/2-3 to Burckhardt) and the Arbeitsgemeinschaft für Pädiatrische Stoffwechselstörungen (APS).

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Authors and Affiliations

  1. Department of Pediatrics, University Medical Center Hamburg-Eppendorf, Martinistrasse 52, 20246, Hamburg, Germany

    Franziska Stellmer, Britta Keyser, Sabrina Jabs, Zoltan Lukacs, Kurt Ullrich, Thomas Braulke & Chris Mühlhausen

  2. Center of Physiology and Pathophysiology, Georg-August-University, Humboldtallee 23, 37073, Göttingen, Germany

    Birgitta C. Burckhardt & Gerhard Burckhardt

  3. Institute of Anatomy and Cell Biology, University of Würzburg, Koellikerstrasse 6, 97070, Würzburg, Germany

    Hermann Koepsell

  4. Department of Clinical Chemistry, University Medical Center Hamburg-Eppendorf, Martinistrasse 52, 20246, Hamburg, Germany

    Thomas Streichert & Zoltan Lukacs

  5. Department of Neuropathology, University Medical Center Hamburg-Eppendorf, Martinistrasse 52, 20246, Hamburg, Germany

    Markus Glatzel

  6. Institute of Organic Chemistry, University of Hamburg, Martin-Luther-King-Platz 6, 20146, Hamburg, Germany

    Joachim Thiem & Wilhelm Herdering

  7. Department of Pediatrics, Oregon Health and Science University, 707 SW Gaines Road, Portland, OR, 97239, USA

    David M. Koeller

  8. Department of Pediatrics, University of Colorado Health Sciences Center, P.O. Box C233, 4200 East Ninth Avenue, Denver, CO, 80262, USA

    Stephen I. Goodman

Authors
  1. Franziska Stellmer
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  2. Britta Keyser
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  3. Birgitta C. Burckhardt
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  4. Hermann Koepsell
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  5. Thomas Streichert
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  6. Markus Glatzel
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  7. Sabrina Jabs
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  8. Joachim Thiem
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  9. Wilhelm Herdering
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  10. David M. Koeller
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  11. Stephen I. Goodman
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  12. Zoltan Lukacs
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  13. Kurt Ullrich
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  14. Gerhard Burckhardt
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  15. Thomas Braulke
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  16. Chris Mühlhausen
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Corresponding author

Correspondence to Chris Mühlhausen.

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Supplementary online Fig. 1

Co-localization of NaDC3 and Kcc4 cotransporter. Cryosections of kidney tissue of 42 day-old Gcdh−/−mice were simultaneously incubated with antibodies against NaDC3 (brown) and Kcc4 (red). Additionally, the Kcc4 localization was visualized by immunofluorescence microscopy. Arrows and arrowheads indicate basolateral and apical membranes, respectively (JPG 183 kb)

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Stellmer, F., Keyser, B., Burckhardt, B.C. et al. 3-Hydroxyglutaric acid is transported via the sodium-dependent dicarboxylate transporter NaDC3. J Mol Med 85, 763–770 (2007). https://doi.org/10.1007/s00109-007-0174-5

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  • DOI: https://doi.org/10.1007/s00109-007-0174-5

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