title: Detecting Cytosolic Peptide Delivery with the GFP Complementation Assay in the Low Micromolar Range.
abstract: Transfection of cells with a plasmid encoding for the first ten strands of the GFP protein (GFP1-10) provides the means to detect cytosolic peptide import at low micromolar concentrations. Cytosolic import of the eleventh strand of the GFP protein either by electroporation or by cell-penetrating peptide-mediated import leads to formation of the full-length GFP protein and fluorescence. An increase in sensitivity is achieved through structural modifications of the peptide and the expression of GFP1-10 as a fusion protein with mCherry.
authors.affiliation: Department of Biochemistry (286), Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, Geert Grooteplein 28, 6525 GA Nijmegen (The Netherlands).
authors.forename: Samuel
authors.initials: S
authors.lastname: Schmidt
authors.affiliation: Department of Biochemistry (286), Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, Geert Grooteplein 28, 6525 GA Nijmegen (The Netherlands).
authors.forename: Merel J W
authors.initials: MJ
authors.lastname: Adjobo-Hermans
authors.affiliation: Department of Biochemistry (286), Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, Geert Grooteplein 28, 6525 GA Nijmegen (The Netherlands).
authors.forename: Rike
authors.initials: R
authors.lastname: Wallbrecher
authors.affiliation: Department of Biochemistry (286), Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, Geert Grooteplein 28, 6525 GA Nijmegen (The Netherlands).|Present address: Department of Biochemistry, University of Zurich, Winterthurerstrasse 190, 8057 Zurich (Switzerland).
authors.forename: Wouter P R
authors.initials: WP
authors.lastname: Verdurmen
authors.affiliation: Department of Biochemistry (286), Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, Geert Grooteplein 28, 6525 GA Nijmegen (The Netherlands).
authors.forename: Petra H M
authors.initials: PH
authors.lastname: Bovée-Geurts
authors.affiliation: Department of Biochemistry (286), Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, Geert Grooteplein 28, 6525 GA Nijmegen (The Netherlands).
authors.forename: Jenny
authors.initials: J
authors.lastname: van Oostrum
authors.affiliation: Roche Pharmaceutical Research and Early Development, Roche Innovation Center New York, 430 East 29th Street, 10016 New York, NY (USA).
authors.forename: Francesca
authors.initials: F
authors.lastname: Milletti
authors.affiliation: Roche Pharmaceutical Research and Early Development, Roche Innovation Center Basel, F. Hoffmann-La Roche Ltd, Grenzacherstrasse 124, 4070 Basel (Switzerland).
authors.forename: Thilo
authors.initials: T
authors.lastname: Enderle
authors.affiliation: Department of Biochemistry (286), Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, Geert Grooteplein 28, 6525 GA Nijmegen (The Netherlands). roland.brock@radboudumc.nl.
authors.forename: Roland
authors.initials: R
authors.lastname: Brock
mesh_terms: D057846:Cell-Penetrating Peptides / Q000032:analysis* / Q000378:metabolism; D003600:Cytosol / Q000737:chemistry / Q000378:metabolism*; D005453:Fluorescence; D049452:Green Fluorescent Proteins / Q000737:chemistry / Q000378:metabolism*; D057809:HEK293 Cells; D006801:Humans
chemical_list: D057846:Cell-Penetrating Peptides; D049452:Green Fluorescent Proteins
keywords: cell uptake; cell-penetrating peptides; drug delivery; fragment complementation assays; lipid membranes