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Wiley, Angewandte Chemie, 50(127), p. 15320-15323, 2015

DOI: 10.1002/ange.201505913

Wiley, Angewandte Chemie International Edition, 50(54), p. 15105-15108, 2015

DOI: 10.1002/anie.201505913

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Detecting Cytosolic Peptide Delivery with the GFP Complementation Assay in the Low Micromolar Range

Journal article published in 2015 by Samuel Schmidt, Merel J. W. Adjobo Hermans ORCID, Rike Wallbrecher, Wouter P. R. Verdurmen ORCID, Petra H. M. Bovée Geurts, Jenny van Oostrum, Francesca Milletti, Thilo Enderle, Roland Brock
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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.