title: Chemolabile cellular microarrays for screening small molecules and peptides.
abstract: Microarrays that mediate the uptake of small molecules into living cells are described. Tissue culture cells were seeded onto glass substrates functionalized locally with fluorescently labelled test substances. In order to enable a localized transfer of substances after contact of cells with the substrate, substances were immobilized on the surface either by non-covalent interactions or chemolabile linker groups. These chemolabile linker groups were incorporated into covalently immobilized compounds. Different ester linkages were evaluated as chemolabile linker groups. As model compounds, esters of the carboxy group of a cysteine with the hydroxy groups of carboxyfluorescein-labelled serine amide and tyrosine amide residues or the thiol group of another fluorescein-labelled cysteine amide were generated. Covalent immobilization occurred on maleimide-functionalized glass cover slips. The surface functionalization and release kinetics were assessed by confocal laser scanning microscopy. The fastest release was obtained for the phenolic tyrosine ester. Alternatively, fluorescently labelled peptides were immobilized by non-covalent interactions on glass and on a hydrogel matrix. In order to increase the efficiency of cellular uptake, peptides were N-terminally extended with a cell-penetrating peptide. Uptake of these peptides into cells was confined to the functionalized spots, and was specific for peptides extended with the cell-penetrating peptide.
authors.affiliation: Institute for Cell Biology, University of Tübingen, Tübingen, Germany.
authors.forename: Antje
authors.initials: A
authors.lastname: Hoff
authors.forename: Thomas
authors.initials: T
authors.lastname: André
authors.forename: Rainer
authors.initials: R
authors.lastname: Fischer
authors.forename: Söhnke
authors.initials: S
authors.lastname: Voss
authors.forename: Michael
authors.initials: M
authors.lastname: Hulko
authors.forename: Udo
authors.initials: U
authors.lastname: Marquardt
authors.forename: Karl-Heinz
authors.initials: KH
authors.lastname: Wiesmüller
authors.forename: Roland
authors.initials: R
authors.lastname: Brock
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002448:Cell Adhesion / Q000502:physiology; D002460:Cell Line; D007202:Indicators and Reagents; D018613:Microscopy, Confocal; D008958:Models, Molecular; D008969:Molecular Sequence Data; D019151:Peptide Library; D010455:Peptides / Q000138:chemical synthesis* / Q000737:chemistry; D040081:Protein Array Analysis; D011487:Protein Conformation
chemical_list: D007202:Indicators and Reagents; D019151:Peptide Library; D010455:Peptides