title: Peptide transporter isoforms are discriminated by the fluorophore-conjugated dipeptides β-Ala- and d-Ala-Lys-N-7-amino-4-methylcoumarin-3-acetic acid.
abstract: Peptide transporters of the SLC15 family are classified by structure and function into PEPT1 (low-affinity/high-capacity) and PEPT2 (high-affinity/low-capacity) isoforms. Despite the differences in kinetics, both transporter isoforms are reckoned to transport essentially all possible di- and tripeptides. We here report that the fluorophore-conjugated dipeptide derivatives β-Ala-Lys-N-7-amino-4-methylcoumarin-3-acetic acid (β-AK-AMCA) and d-Ala-Lys-N-7-amino-4-methylcoumarin-3-acetic acid (d-AK-AMCA) are transported by distinct PEPT isoforms in a species-specific manner. Transport of the fluorophore peptides was studied (1) in vitro after heterologous expression in Xenopus oocytes of PEPT1 and PEPT2 isoforms from different vertebrate species and of PEPT1 and PEPT2 transporters from Caenorhabditis elegans by using electrophysiological and fluorescence methods and (2) in vivo in C. elegans by using fluorescence methods. Our results indicate that both substrates are transported by the vertebrate "renal-type" and the C. elegans "intestinal-type" peptide transporter only. A systematic analysis among species finds four predicted amino acid residues along the sequence that may account for the substrate uptake differences observed between the vertebrate PEPT1/nematode PEPT2 and the vertebrate PEPT2/nematode PEPT1 subtype. This selectivity on basis of isoforms and species may be helpful in better defining the structure-function determinants of the proteins of the SLC15 family.
authors.affiliation: ZIEL Research Center of Nutrition and Food Sciences, Abteilung Biochemie, Technische Universität München, Gregor-Mendel-Str. 2, Freising, D-85350, Germany.
authors.forename: Gabor
authors.initials: G
authors.lastname: Kottra
authors.affiliation: ZIEL Research Center of Nutrition and Food Sciences, Abteilung Biochemie, Technische Universität München, Gregor-Mendel-Str. 2, Freising, D-85350, Germany.
authors.forename: Britta
authors.initials: B
authors.lastname: Spanier
authors.affiliation: Laboratory of General Physiology, Department of Biological and Environmental Sciences and Technologies, University of Salento, via Provinciale Lecce-Monteroni, Lecce, I-73100, Italy.
authors.forename: Tiziano
authors.initials: T
authors.lastname: Verri
authors.affiliation: ZIEL Research Center of Nutrition and Food Sciences, Abteilung Biochemie, Technische Universität München, Gregor-Mendel-Str. 2, Freising, D-85350, Germany.
authors.forename: Hannelore
authors.initials: H
authors.lastname: Daniel
keywords: Caenorhabditis elegans; PEPT1; PEPT2; di‐ and tripeptide; substrate specificity; transporter
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