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SPAK-Sensitive Regulation of Glucose Transporter SGLT1

The Journal of Membrane Biology volume 247, pages 1191–1197 (2014)Cite this article

Abstract

The WNK-dependent STE20/SPS1-related proline/alanine-rich kinase SPAK is a powerful regulator of ion transport. The study explored whether SPAK similarly regulates nutrient transporters, such as the Na+-coupled glucose transporter SGLT1 (SLC5A1). To this end, SGLT1 was expressed in Xenopus oocytes with or without additional expression of wild-type SPAK, constitutively active T233ESPAK, WNK-insensitive T233ASPAK or catalytically inactive D212ASPAK, and electrogenic glucose transport determined by dual-electrode voltage-clamp experiments. Moreover, Ussing chamber was employed to determine the electrogenic glucose transport in intestine from wild-type mice (spak wt/wt) and from gene-targeted mice carrying WNK-insensitive SPAK (spak tg/tg). In SGLT1-expressing oocytes, but not in water-injected oocytes, the glucose-dependent current (I g) was significantly decreased following coexpression of wild-type SPAK and T233ESPAK, but not by coexpression of T233ASPAK or D212ASPAK. Kinetic analysis revealed that SPAK decreased maximal I g without significantly modifying the glucose concentration required for halfmaximal I g (K m). According to the chemiluminescence experiments, wild-type SPAK but not D212ASPAK decreased SGLT1 protein abundance in the cell membrane. Inhibition of SGLT1 insertion by brefeldin A (5 μM) resulted in a decline of I g, which was similar in the absence and presence of SPAK, suggesting that SPAK did not accelerate the retrieval of SGLT1 protein from the cell membrane but rather down-regulated carrier insertion into the cell membrane. Intestinal electrogenic glucose transport was significantly lower in spak wt/wt than in spak tg/tg mice. In conclusion, SPAK is a powerful negative regulator of SGLT1 protein abundance in the cell membrane and thus of electrogenic glucose transport.

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Acknowledgments

The authors acknowledge the meticulous preparation of the manuscript by Sari Rübe and technical support by Elfriede Faber. This study was supported by the Deutsche Forschungsgemeinschaft, GRK 1302, SFB 773 B4/A1, La 315/13-3.

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  1. Department of Physiology, University of Tübingen, Gmelinstr. 5, 72076, Tübingen, Germany

    Bernat Elvira, Maria Blecua, Dong Luo, Wenting Yang, Ekaterina Shumilina & Florian Lang

  2. Institute of Physiology and Zurich Center for Integrative Human Physiology (ZIHP), University of Zurich, Winterthurerstrasse 190, 8057, Zurich, Switzerland

    Carlos Munoz

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  1. Bernat Elvira
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  2. Maria Blecua
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  3. Dong Luo
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Correspondence to Florian Lang.

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Bernat Elvira and Maria Blecua contributed equally and thus share first authorship.

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Elvira, B., Blecua, M., Luo, D. et al. SPAK-Sensitive Regulation of Glucose Transporter SGLT1. J Membrane Biol 247, 1191–1197 (2014). https://doi.org/10.1007/s00232-014-9719-z

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Keywords

  • Glucose transport
  • Intestine
  • Oocytes
  • Mice
  • WNK