Profiling at mRNA, protein, and metabolite levels reveals alterations in renal amino acid handling and glutathione metabolism in kidney tissue of Pept2-/- mice

Isabelle M. Frey, Isabel Rubio-Aliaga, Anne Siewert, Daniela Sailer, Aleksey Drobyshev, Johannes Beckers, Martin Hrabé De Angelis, Julie Aubert, Avner Bar Hen, Oliver Fiehn, Hans M. Eichinger, Hannelore Daniel

Research output: Contribution to journalArticle

48 Scopus citations

Abstract

PEPT2 is an integral membrane protein in the apical membrane of renal epithelial cells that operates as a rheogenic transporter for di- and tripeptides and structurally related drugs. Its prime role is thought to be the reabsorption of filtered di- and tripeptides contributing to amino acid homeostasis. To elucidate the role of PEPT2 in renal amino acid metabolism we submitted kidney tissues of wild-type and a Pept2-/- mouse line to a comprehensive transcriptome, proteome and metabolome profiling and analyzed urinary amino acids and dipeptides. cDNA microarray analysis identified 147 differentially expressed transcripts in transporter-deficient animals, and proteome analysis by 2D-PAGE and MALDI-TOF-MS identified 37 differentially expressed proteins. Metabolite profiling by GC-MS revealed predominantly altered concentrations of amino acids and derivatives. Urinary excretion of amino acids demonstrated increased glycine and cysteine/cystine concentrations and dipeptides in urine were assessed by amino acid analysis of urine samples before and after in vitro dipeptidase digestion. Dipeptides constituted a noticeable fraction of urinary amino acids in Pept2-/- animals, only, and dipeptide-bound glycine and cystine were selectively increased in Pept2 -/- urine samples. These findings were confirmed by a drastically increased excretion of cysteinyl-glycine (cys-gly). Urinary loss of cys-gly together with lower concentrations of cysteine, glycine, and oxoproline in kidney tissue and altered expression of mRNA and proteins involved in glutathione (GSH) metabolism suggests that PEPT2 is predominantly a system for reabsorption of cys-gly originating from GSH break-down, thus contributing to resynthesis of GSH.

Original languageEnglish (US)
Pages (from-to)301-310
Number of pages10
JournalPhysiological Genomics
Volume28
Issue number3
DOIs
StatePublished - Feb 12 2007

Keywords

  • Glutathione metabolism
  • Pathway analysis
  • PEPT2
  • Peptide transport

ASJC Scopus subject areas

  • Physiology
  • Genetics

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    Frey, I. M., Rubio-Aliaga, I., Siewert, A., Sailer, D., Drobyshev, A., Beckers, J., De Angelis, M. H., Aubert, J., Hen, A. B., Fiehn, O., Eichinger, H. M., & Daniel, H. (2007). Profiling at mRNA, protein, and metabolite levels reveals alterations in renal amino acid handling and glutathione metabolism in kidney tissue of Pept2-/- mice. Physiological Genomics, 28(3), 301-310. https://doi.org/10.1152/physiolgenomics.00193.2006