title: New transport peptides broaden the horizon of applications for peptidic pharmaceuticals.
abstract: Protein transduction domains (PTDs) have proven to be an invaluable tool to transduce a wide variety of cargo's including peptides across the plasma membrane and into intact tissue. The PTDs are able to deliver biologically active molecules both in vitro and in vivo. This study describes many new polybasic PTDs of which some are just as potent as the PTDs derived from extracellular RNAses or other published PTDs. Large differences in potency became apparent when the PTDs are coupled to particular cargoes. Therefore, the unique characteristic of a PTD may only become apparent when it is selected for a particular application. Rules for optimization of PTDs for particular applications are now emerging and open the way for a new generation of drug delivery agents. Because fixation artifacts and irreversible membrane binding may cause misinterpretation of the amount of internalization of polybasic peptides, we have developed an enzyme transduction assay based on the intracellular loading of a cell permeable substrate. In this assay, a fluorescent signal is generated by internalized enzyme in intact cells and not by membrane-bound or extracellular enzyme.
authors.affiliation: Pepscan Systems B.V., Lelystad, The Netherlands. j.p.m.langedijk@pepscan.nl
authors.forename: J P M
authors.initials: JP
authors.lastname: Langedijk
authors.forename: T
authors.initials: T
authors.lastname: Olijhoek
authors.forename: D
authors.initials: D
authors.lastname: Schut
authors.forename: R
authors.initials: R
authors.lastname: Autar
authors.forename: R H
authors.initials: RH
authors.lastname: Meloen
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism; D002417:Cattle; D002478:Cells, Cultured; D016650:Fluorescein-5-isothiocyanate / Q000737:chemistry / Q000378:metabolism; D005452:Fluoresceins / Q000737:chemistry / Q000378:metabolism; D005456:Fluorescent Dyes / Q000737:chemistry / Q000378:metabolism; D005697:Galactosides / Q000737:chemistry / Q000378:metabolism; D015696:Gene Products, tat / Q000235:genetics / Q000378:metabolism; D006493:Heparin / Q000378:metabolism; D006735:Horseradish Peroxidase / Q000235:genetics / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism; D008967:Molecular Biology / Q000379:methods*; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D010455:Peptides / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D015202:Protein Engineering / Q000379:methods; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D019809:Streptavidin / Q000737:chemistry / Q000378:metabolism; D014759:Viral Envelope Proteins / Q000235:genetics / Q000378:metabolism; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
chemical_list: D002352:Carrier Proteins; C466501:E(rns) protein, classic swine fever virus; D005452:Fluoresceins; D005456:Fluorescent Dyes; C066028:G protein, Bovine respiratory syncytial virus; D005697:Galactosides; D015696:Gene Products, tat; D008562:Membrane Glycoproteins; D010455:Peptides; D014759:Viral Envelope Proteins; C038848:fluorescein-digalactoside; D006493:Heparin; D019809:Streptavidin; D006735:Horseradish Peroxidase; D001616:beta-Galactosidase; D016650:Fluorescein-5-isothiocyanate