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

44 Citations (Scopus)

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

Fingerprint

Dipeptides
Glutathione
cysteinylglycine
Kidney
Amino Acids
Messenger RNA
Proteins
Glycine
Cystine
Urine
Proteome
Cysteine
Metabolome
Matrix-Assisted Laser Desorption-Ionization Mass Spectrometry
Electrophoresis, Gel, Two-Dimensional
Microarray Analysis
Oligonucleotide Array Sequence Analysis
Transcriptome
Digestion
Membrane Proteins

Keywords

  • Glutathione metabolism
  • Pathway analysis
  • PEPT2
  • Peptide transport

ASJC Scopus subject areas

  • Physiology
  • Genetics

Cite this

Profiling at mRNA, protein, and metabolite levels reveals alterations in renal amino acid handling and glutathione metabolism in kidney tissue of Pept2-/- mice. / Frey, Isabelle M.; Rubio-Aliaga, Isabel; Siewert, Anne; Sailer, Daniela; Drobyshev, Aleksey; Beckers, Johannes; De Angelis, Martin Hrabé; Aubert, Julie; Hen, Avner Bar; Fiehn, Oliver; Eichinger, Hans M.; Daniel, Hannelore.

In: Physiological Genomics, Vol. 28, No. 3, 12.02.2007, p. 301-310.

Research output: Contribution to journalArticle

Frey, IM, Rubio-Aliaga, I, Siewert, A, Sailer, D, Drobyshev, A, Beckers, J, De Angelis, MH, Aubert, J, Hen, AB, Fiehn, O, Eichinger, HM & 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, vol. 28, no. 3, pp. 301-310. https://doi.org/10.1152/physiolgenomics.00193.2006
Frey, Isabelle M. ; Rubio-Aliaga, Isabel ; Siewert, Anne ; Sailer, Daniela ; Drobyshev, Aleksey ; Beckers, Johannes ; De Angelis, Martin Hrabé ; Aubert, Julie ; Hen, Avner Bar ; Fiehn, Oliver ; Eichinger, Hans M. ; Daniel, Hannelore. / Profiling at mRNA, protein, and metabolite levels reveals alterations in renal amino acid handling and glutathione metabolism in kidney tissue of Pept2-/- mice. In: Physiological Genomics. 2007 ; Vol. 28, No. 3. pp. 301-310.
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AU - Sailer, Daniela

AU - Drobyshev, Aleksey

AU - Beckers, Johannes

AU - De Angelis, Martin Hrabé

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AU - Fiehn, Oliver

AU - Eichinger, Hans M.

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