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The glycolytic enzymes glyceraldehyde 3-phosphate dehydrogenase and enolase interact with the renal epithelial K+ channel ROMK2 and regulate its function.
- Source :
-
Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology [Cell Physiol Biochem] 2011; Vol. 28 (4), pp. 663-72. Date of Electronic Publication: 2011 Dec 14. - Publication Year :
- 2011
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Abstract
- Background/aims: ROMK channels mediate potassium secretion and regulate NaCl reabsorption in the kidney. The aim was to study the functional implications of the interaction between ROMK2 (Kir1.1b) and two glycolytic enzymes, glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and enolase-α, which were identified as potential regulatory subunits of the channel complex.<br />Methods: We performed a membrane yeast-two-hybrid screen of a human kidney cDNA library with ROMK2 as a bait. Interaction of ROMK2 with GAPDH and enolase was verified using GST pull-down, co-immunoprecipitation, immunohistochemistry and co-expression in Xenopus oocytes.<br />Results: Confocal imaging showed co-localisation of enolase and GAPDH with ROMK2 in the apical membrane of the renal epithelial cells of the thick ascending limb. Over-expression of GAPDH or enolase-α in Xenopus oocytes markedly reduced the amplitude of ROMK2 currents but did not affect the surface expression of the channels. Co-expression of the glycolytically inactive GAPDH mutant C149G did not have any effect on ROMK2 current amplitude.<br />Conclusion: Our results suggest that the glycolytic enzymes GAPDH and enolase are part of the ROMK2 channel supramolecular complex and may serve to couple salt reabsorption in the thick ascending limb of the loop of Henle to the metabolic status of the renal epithelial cells.<br /> (Copyright © 2011 S. Karger AG, Basel.)
- Subjects :
- Amino Acid Substitution
Animals
Glyceraldehyde-3-Phosphate Dehydrogenases genetics
Glyceraldehyde-3-Phosphate Dehydrogenases physiology
HEK293 Cells
Humans
Immunoprecipitation
Kidney enzymology
Kidney metabolism
Oocytes metabolism
Patch-Clamp Techniques
Potassium Channels, Inwardly Rectifying metabolism
Two-Hybrid System Techniques
Xenopus laevis genetics
Glyceraldehyde-3-Phosphate Dehydrogenases metabolism
Phosphopyruvate Hydratase metabolism
Potassium Channels, Inwardly Rectifying physiology
Subjects
Details
- Language :
- English
- ISSN :
- 1421-9778
- Volume :
- 28
- Issue :
- 4
- Database :
- MEDLINE
- Journal :
- Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology
- Publication Type :
- Academic Journal
- Accession number :
- 22178878
- Full Text :
- https://doi.org/10.1159/000335761