title: Tailored Presentation of Carbohydrates on a Coiled Coil-Based Scaffold for Asialoglycoprotein Receptor Targeting.
abstract: The coiled-coil folding motif represents an ideal scaffold for the defined presentation of ligands due to the possibility of positioning them at specific distances along the axis. We created a coiled-coil glycopeptide library to characterize the distances between the carbohydrate-binding sites of the asialoglycoprotein receptors (ASGPR) on hepatocytes. The components of the glycopeptide library vary for the number of displayed ligands (galactose), their position on the peptide sequence, and the space between peptide backbone and carbohydrate. We determined the binding of the glycopeptides to the hepatocytes, and we established the optimal distance and orientation of the galactose moieties for interaction with the ASGPR using flow cytometry. We confirmed that the binding occurs through endocytosis mediated by ASGPR via inhibition studies with cytochalasin D; fluorescence microscopy studies display the uptake of the carrier peptides inside the cell. Thus, this study demonstrates that the coiled-coil motif can be used as reliable scaffold for the rational presentation of ligands.
authors.affiliation: Institute of Chemistry and Biochemistry, Freie Universität Berlin , Takustrasse 3, 14195 Berlin, Germany.
authors.forename: Elsa
authors.initials: E
authors.lastname: Zacco
authors.affiliation: Department of Biomolecular Systems, Max Planck Institute of Colloids and Interfaces , Am Mühlenberg 1, 14424 Potsdam, Germany.|Institute of Chemistry and Biochemistry, Freie Universität Berlin , Arnimallee 22, 14195 Berlin, Germany.
authors.forename: Julia
authors.initials: J
authors.lastname: Hütter
authors.affiliation: Institute of Chemistry and Biochemistry, Freie Universität Berlin , Takustrasse 3, 14195 Berlin, Germany.
authors.forename: Jason L
authors.initials: JL
authors.lastname: Heier
authors.affiliation: Institute of Chemistry and Biochemistry, Freie Universität Berlin , Takustrasse 3, 14195 Berlin, Germany.|Institute of Pharmacy, Freie Universität Berlin , Königin-Luisestrasse 2, 14195 Berlin, Germany.
authors.forename: Jérémie
authors.initials: J
authors.lastname: Mortier
authors.affiliation: Department of Biomolecular Systems, Max Planck Institute of Colloids and Interfaces , Am Mühlenberg 1, 14424 Potsdam, Germany.|Institute of Chemistry and Biochemistry, Freie Universität Berlin , Arnimallee 22, 14195 Berlin, Germany.
authors.forename: Peter H
authors.initials: PH
authors.lastname: Seeberger
authors.affiliation: Department of Biomolecular Systems, Max Planck Institute of Colloids and Interfaces , Am Mühlenberg 1, 14424 Potsdam, Germany.|Institute of Chemistry and Biochemistry, Freie Universität Berlin , Arnimallee 22, 14195 Berlin, Germany.
authors.forename: Bernd
authors.initials: B
authors.lastname: Lepenies
authors.affiliation: Institute of Chemistry and Biochemistry, Freie Universität Berlin , Takustrasse 3, 14195 Berlin, Germany.
authors.forename: Beate
authors.initials: B
authors.lastname: Koksch
mesh_terms: D000595:Amino Acid Sequence; D037263:Asialoglycoprotein Receptor / Q000378:metabolism*; D001665:Binding Sites; D005690:Galactose / Q000737:chemistry / Q000378:metabolism*; D006020:Glycopeptides / Q000737:chemistry / Q000378:metabolism*; D056945:Hep G2 Cells; D022781:Hepatocytes / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D008024:Ligands; D008969:Molecular Sequence Data; D019151:Peptide Library; D011485:Protein Binding; D017433:Protein Structure, Secondary
chemical_list: D037263:Asialoglycoprotein Receptor; D006020:Glycopeptides; D008024:Ligands; D019151:Peptide Library; D005690:Galactose