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Renal Localization and Characterization of Monocarboxylate Transporter Family Member Slc16a14
Author(s) -
Knöpfel Thomas,
Atanassoff Alexander,
Kampik Nicole,
Hernando Nati,
Wagner Carsten,
Biber Jürg
Publication year - 2015
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.29.1_supplement.969.13
Subject(s) - xenopus , monocarboxylate transporter , solute carrier family , nephron , symporter , biology , chemistry , kidney , microbiology and biotechnology , transporter , biochemistry , endocrinology , gene
Monocarboxylate transporter 14 (MCT14) is an orphan member of the solute carrier family 16 for monocarboxylates, thyroid hormone and aromatic amino acids. The kidney shows the highest abundance of MCT14 mRNA, however, the renal localization, function and regulation are poorly understood. The aim was to localize MCT14 in murine kidney and address the question of its possible function. To this end an antibody was raised against a peptide located at the extracellular side of MCT14 and its specifity was confirmed by peptide protection and heterologous expression of MCT14 in Xenopus laevis oocytes. Colocalization experiments with markers for different nephron segments indicate that MCT14 is localized to the apical membrane of the thick ascending limb of the loop of Henle. Members of the Slc16 family showed changes in protein and mRNA abundance in kidney due to metabolic acidosis. MCT14 itself showed an upregulation under acidotic conditions in both mRNA and protein levels, but the expression pattern in the kidney was not altered. Uptake experiments in Xenopus laevis oocytes expressing MCT14 suggest that MCT14 does not transport lactate or pyruvate unlike the classical monocarboxylate transporters of the Slc16 family. To identify the substrate of MCT14 we used a mass spectrometry based approach in order to cover many substrates. Therefore Xenopus laevis oocytes expressing MCT14 and corresponding negative controls were incubated in mouse serum. Intracellular metabolites were then analyzed by mass spectrometry in the Functional Genomics Center Zurich. Major substrate candidates identified by mass spectrometry are currently being examined to confirm whether they are substrates of MCT14. Funded by the Swiss National Science Foundation.