Transcellular transfer of active HSV-1 thymidine kinase mediated by an 11-amino-acid peptide from HIV-1 Tat

Tasciotti, Ennio; Zoppé, Monica; Giacca, Mauro

doi:10.1038/sj.cgt.7700526

Cited by 47 publications

(30 citation statements)

References 51 publications

(46 reference statements)

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“…[4][5][6][7][8][9] PTDs can even deliver bulky molecular cargos (>100 kDa) into a wide variety of cell types. [10][11][12][13] However, to use PTDs as effective intracellular drug delivery carriers with clinical applications, it is necessary to create novel PTDs with greater protein-transduction potency than exists naturally.…”

Section: -3)mentioning

confidence: 99%

Creation of Novel Protein Transduction Domain (PTD) Mutants by a Phage Display-Based High-Throughput Screening System

Mukai

Sugita

Yamato

et al. 2006

Biological & Pharmaceutical Bulletin

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Recent advances in proteomics have allowed a number of refractory diseases, such as cancer and neurodegenerative disorders, to be studied at the molecular level. The main causative factor of such disease states is often associated with intracellular organelles or particular subcellular proteins. Thus, the intracellular organelles, proteins or genes might constitute the therapeutic target. Recently, it was discovered that certain peptides, referred to as protein transduction domains (PTDs), can penetrate cells accompanied by a large molecular cargo. Considerable research effort is currently focused on utilizing PTDs as peptide-based carriers for intracellular drug delivery. 1-3)Tat peptide, derived from the HIV-1, and Antennapedia peptide, derived from Drosophila Antennapedia homeotic transcription factor, are well known PTDs that have been tested as drug delivery carriers for various disease models. [4][5][6][7][8][9] PTDs can even deliver bulky molecular cargos (>100 kDa) into a wide variety of cell types. [10][11][12][13] However, to use PTDs as effective intracellular drug delivery carriers with clinical applications, it is necessary to create novel PTDs with greater protein-transduction potency than exists naturally.An attempt to create a novel PTD by modification of the peptide structure has already been reported. 14,15) However, because it is difficult to predict the transduction activity of the peptide based on structural information alone, novel peptides must be generated by introducing amino acid substitutions and then the effects determined by trial and error. Recently, we have successfully generated a technology for creating novel muteins (mutant proteins) that have non-native functions using a phage display system.16) This prompted us to apply phage display technology to screen for novel PTDs.The phage display system is a protein selection methodology in which a library of mutant proteins or peptides can be screened and the desired molecules easily identified by linking DNA information (genotype) with phenotype (protein expression). [16][17][18][19][20] By applying this methodology, novel PTDs can be selected such as those transduced into the cell by a different mechanism or those with tissue/cell specificity. In general, the phage display system is used to isolate antibody and peptide ligands using an affinity selection step to target the desired molecules. However, for the discovery of PTDs it is necessary to construct a screening method to select clones that are transduced into the cell rather than simply selecting those that bind to the cell surface. We designed a highthroughput screening method to isolate effective PTDs by fusing PTD with Protein Synthesis Inhibitory Factor (PSIF).21) Here, we used our methodology to identify novel Tat mutants with greater transduction potency than wild-type Tat PTD. ; 7-6-8 Saito-Asagi, Ibaraki, Osaka 567-0085, Japan: and b Department of Biopharmaceutics, Graduate School of Pharmaceutical Sciences, Osaka University; 1-6 Yamadaoka, Suita, Osaka 565-0871, Ja...

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Section: -3)mentioning

confidence: 99%

Creation of Novel Protein Transduction Domain (PTD) Mutants by a Phage Display-Based High-Throughput Screening System

Mukai

Sugita

Yamato

et al. 2006

Biological & Pharmaceutical Bulletin

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“…Limited studies have shown that PTD is conferred with transcellular property [125,141]. Indeed, this behavior is extremely valuable in delivering therapeutic proteins to surrounding cells.…”

Section: Transcellular Property Of Ptdmentioning

confidence: 99%

The taming of the cell penetrating domain of the HIV Tat: Myths and realities

Chauhan

Tikoo²,

Kapur

et al. 2007

Journal of Controlled Release

156

136

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Protein transduction with cell penetrating peptides over the past several years has been shown to be an effective way of delivering proteins in vitro and now several reports have also shown valuable in vivo applications in correcting disease states. An impressive bioinspired phenomenon of crossing biological barriers came from HIV transactivator Tat protein. Specifically, the protein transduction domain of HIV-Tat has been shown to be a potent pleiotropic peptide in protein delivery. Various approaches such as molecular modeling, arginine guanidinium head group structural strategy, multimerization of PTD sequence and phage display system have been applied for taming of the PTD. This has resulted in identification of PTD variants which are efficient in cell membrane penetration and cytoplasmic delivery. Inspite of these state of the art technologies, the dilemma of low protein transduction efficiency and target specific delivery of PTD fusion proteins remains unsolved. Moreover, some misconceptions about PTD of Tat in the literature require considerations. We have assembled critical information on secretory, plasma membrane penetration and transcellular properties of Tat and PTD using molecular analysis and available experimental evidences.

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“…Thus, development of this approach was an important step in the establishment of TK suicide gene therapy. 236 The attachment of TATp to a water-soluble synthetic macromolecule, N-(2-hydroxypropyl)methacrylamide copolymer, resulted in the cytoplasmic and nuclear delivery of the conjugate via nonendocytotic and concentration-independent processes, as opposed to conjugates without TATp, which accumulated in only endocytotic vesicles. Furthermore, the TATp-polymer-bound anticancer drug doxorubicin was delivered inside the cytoplasm, providing the possibility for the development of polymer-based systems for the cytoplasmic delivery of therapeutic molecules.…”

Section: Delivery By Cppsmentioning

confidence: 99%