title: Creation of novel Protein Transduction Domain (PTD) mutants by a phage display-based high-throughput screening system.
abstract: Significant research effort is currently focused on Protein Transduction Domains (PTDs) as potential intracellular drug delivery carriers. However, the application of this technology is limited because the transduction efficiencies are often insufficient for therapeutic purposes, even using HIV-1 Tat peptide. Here we describe a high-throughput screening method based on a phage display system for isolating novel PTDs with improved cell penetration activity. The screening method involves using protein synthesis inhibitory factor (PSIF) as cargo of PTD. Using this method, several Tat-PTD mutants of superior cell-penetrating activity were isolated. Interestingly, the amino acid sequence of the PTD mutants contained some characteristic residues, such as proline. Thus, our screening method may prove useful in determining the relationship between protein transduction and amino acid sequence.
authors.affiliation: Laboratory of Pharmaceutical Proteomics, National Institute of Biomedical Innovation, Japan.
authors.forename: Yohei
authors.initials: Y
authors.lastname: Mukai
authors.forename: Toshiki
authors.initials: T
authors.lastname: Sugita
authors.forename: Tomoko
authors.initials: T
authors.lastname: Yamato
authors.forename: Natsue
authors.initials: N
authors.lastname: Yamanada
authors.forename: Hiroko
authors.initials: H
authors.lastname: Shibata
authors.forename: Sunao
authors.initials: S
authors.lastname: Imai
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Abe
authors.forename: Kazuya
authors.initials: K
authors.lastname: Nagano
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Nomura
authors.forename: Yasuo
authors.initials: Y
authors.lastname: Tsutsumi
authors.forename: Haruhiko
authors.initials: H
authors.lastname: Kamada
authors.forename: Shinsaku
authors.initials: S
authors.lastname: Nakagawa
authors.forename: Shin-ichi
authors.initials: S
authors.lastname: Tsunoda
mesh_terms: D001435:Bacteriophages / Q000235:genetics*; D001483:Base Sequence; D017931:DNA Primers; D006367:HeLa Cells; D006801:Humans; D008856:Microscopy, Fluorescence; D009154:Mutation; D011506:Proteins / Q000737:chemistry / Q000235:genetics*; D014161:Transduction, Genetic
chemical_list: D017931:DNA Primers; D011506:Proteins