Organelle-targeted delivery of biological macromolecules using the protein transduction domain: potential applications for Peptide aptamer delivery into the nucleus.
Extensive effort is currently being expended on the innovative design and engineering of new molecular carrier systems for the organelle-targeted delivery of biological cargoes (e.g., peptide aptamers or biological proteins) as tools in cell biology and for developing novel therapeutic approaches. Although cell-permeable Tat peptides are useful carriers for delivering biological molecules into the cell, much internalized Tat-fused cargo is trapped within macropinosomes and thus not delivered into organelles. Here, we devised a novel intracellular targeting technique to deliver Tat-fused cargo into the nucleus using an endosome-disruptive peptide (hemagglutinin-2 subunit) and a nuclear localization signal peptide. We show for the first time that Tat-conjugated peptide aptamers can be selectively delivered to the nucleus by using combined hemagglutinin-2 subunit and nuclear localization signal peptides. This nuclear targeting technique resulted in marked enhancement of the cytostatic activity of a Tat-fused p53-derived peptide aptamer against human MDM2 (mouse double minute 2) that inhibits p53-MDM2 binding. Thus, our technique provides a unique methodology for the development of novel therapeutic approaches based on intracellular targeting.
Yoshikawa
Tomoaki
T
Laboratory of Pharmaceutical Proteomics, National Institute of Biomedical Innovation, 7-6-8 Saito-Asagi, Ibaraki, Osaka 567-0085, Japan.
Sugita
Toshiki
T
Mukai
Yohei
Y
Yamanada
Natsue
N
Nagano
Kazuya
K
Nabeshi
Hiromi
H
Yoshioka
Yasuo
Y
Nakagawa
Shinsaku
S
Abe
Yasuhiro
Y
Kamada
Haruhiko
H
Tsunoda
Shin-ichi
S
Tsutsumi
Yasuo
Y
D052158:Aptamers, Peptide / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D002460:Cell Line; D002467:Cell Nucleus / Q000737:chemistry / Q000378:metabolism*; D002470:Cell Survival / Q000187:drug effects; D016503:Drug Delivery Systems / Q000379:methods*; D015195:Drug Design; D015696:Gene Products, tat / Q000737:chemistry; D015497:HIV-1 / Q000737:chemistry; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D019913:Nuclear Localization Signals; D010455:Peptides / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology*; D017434:Protein Structure, Tertiary; D021381:Protein Transport / Q000187:drug effects / Q000502:physiology; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D017422:Sequence Analysis, DNA; D013997:Time Factors
D052158:Aptamers, Peptide; D015696:Gene Products, tat; D019913:Nuclear Localization Signals; D010455:Peptides; D011993:Recombinant Fusion Proteins