title: A Biotin Ligase-Based Assay for the Quantification of the Cytosolic Delivery of Therapeutic Proteins.
abstract: The efficient delivery of external proteins from the external milieu to the cytosol of mammalian cells has great potential for both scientific investigations and future therapies. However, when assessing the cellular uptake of proteins, it is often difficult to distinguish between proteins that are stuck in the endosomes and those that have escaped into the cytosol. Here, we describe a method employing the prokaryotic enzyme biotin ligase that overcomes this problem and which can be employed for a highly sensitive quantification of cytosolic protein delivery.
authors.affiliation: Department of Biochemistry, University of Zurich, Winterthurerstr. 190, 8057, Zurich, Switzerland.
authors.forename: Wouter P R
authors.initials: WPR
authors.lastname: Verdurmen
authors.affiliation: Department of Biochemistry, University of Zurich, Winterthurerstr. 190, 8057, Zurich, Switzerland.
authors.forename: Marigona
authors.initials: M
authors.lastname: Mazlami
authors.affiliation: Department of Biochemistry, University of Zurich, Winterthurerstr. 190, 8057, Zurich, Switzerland. Plueckthun@bioc.uzh.ch.
authors.forename: Andreas
authors.initials: A
authors.lastname: Plückthun
mesh_terms: D019731:Carbon-Nitrogen Ligases / Q000235:genetics / Q000378:metabolism*; D003600:Cytosol / Q000378:metabolism*; D011992:Endosomes / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D015202:Protein Engineering; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D019809:Streptavidin / Q000378:metabolism*
chemical_list: D011993:Recombinant Fusion Proteins; D019809:Streptavidin; D019731:Carbon-Nitrogen Ligases
keywords: Biotin; Biotin ligase; Cellular internalization; Cytosolic protein delivery; Protein engineering; Streptavidin; Western blot