Creation of novel Protein Transduction Domain (PTD) mutants by a phage display-based high-throughput screening system.
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.
Mukai
Yohei
Y
Laboratory of Pharmaceutical Proteomics, National Institute of Biomedical Innovation, Japan.
Sugita
Toshiki
T
Yamato
Tomoko
T
Yamanada
Natsue
N
Shibata
Hiroko
H
Imai
Sunao
S
Abe
Yasuhiro
Y
Nagano
Kazuya
K
Nomura
Tetsuya
T
Tsutsumi
Yasuo
Y
Kamada
Haruhiko
H
Nakagawa
Shinsaku
S
Tsunoda
Shin-ichi
S
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
D017931:DNA Primers; D011506:Proteins