Synthetic protein transduction domains: enhanced transduction potential in vitro and in vivo.

The protein transduction domain (PTD) embedded in the HIV TAT protein (amino acids 47-57) has been shown to successfully mediate the introduction of heterologous peptides and proteins in excess of Mr 100,000 into mammalian cells in vitro and in vivo. We report here that the modeled structure of the TAT PTD is a strong amphipathic helix. On the basis of this information, we synthesized a series of synthetic PTDs that strengthen the alpha-helical content and optimize the placement of arginine residues. Several PTD peptides possessed significantly enhanced protein transduction potential compared with TAT in vitro and in vivo. These optimized PTDs have the potential to deliver both existing and novel anticancer therapeutics.

[1]  K. Hruska,et al.  Protein transduction: unrestricted delivery into all cells? , 2000, Trends in cell biology.

[2]  S. Schwarze,et al.  In vivo protein transduction: delivery of a biologically active protein into the mouse. , 1999, Science.

[3]  S. Dowdy,et al.  Transduced p16INK4a peptides inhibit hypophosphorylation of the retinoblastoma protein and cell cycle progression prior to activation of Cdk2 complexes in late G1. , 1999, Cancer research.

[4]  J. Hawiger,et al.  Noninvasive intracellular delivery of functional peptides and proteins. , 1999, Current opinion in chemical biology.

[5]  Natalie A. Lissy,et al.  Transduction of full-length TAT fusion proteins into mammalian cells: TAT-p27Kip1 induces cell migration , 1998, Nature Medicine.

[6]  R. Egleton,et al.  Bioavailability and Transport of Peptides and Peptide Drugs into the Brain , 1997, Peptides.

[7]  Priscille Brodin,et al.  A Truncated HIV-1 Tat Protein Basic Domain Rapidly Translocates through the Plasma Membrane and Accumulates in the Cell Nucleus* , 1997, The Journal of Biological Chemistry.

[8]  I. Luque,et al.  Structure-based thermodynamic scale of alpha-helix propensities in amino acids. , 1996, Biochemistry.

[9]  R. Srinivasan,et al.  LINUS: A hierarchic procedure to predict the fold of a protein , 1995, Proteins.

[10]  K. Sharp,et al.  Protein folding and association: Insights from the interfacial and thermodynamic properties of hydrocarbons , 1991, Proteins.

[11]  E. Vivés,et al.  Lethal neurotoxicity in mice of the basic domains of HIV and SIV Rev proteins Study of these regions by circular dichroism , 1991, FEBS letters.

[12]  E. Vivés,et al.  Activating region of HIV-1 Tat protein: vacuum UV circular dichroism and energy minimization. , 1991, Biochemistry.

[13]  D. Hudson,et al.  Analysis of arginine-rich peptides from the HIV Tat protein reveals unusual features of RNA-protein recognition. , 1991, Genes & development.

[14]  W. Scheld,et al.  Drug delivery to the central nervous system: general principles and relevance to therapy for infections of the central nervous system. , 1989, Reviews of infectious diseases.

[15]  Carl O. Pabo,et al.  Cellular uptake of the tat protein from human immunodeficiency virus , 1988, Cell.

[16]  Maurice Green,et al.  Autonomous functional domains of chemically synthesized human immunodeficiency virus tat trans-activator protein , 1988, Cell.

[17]  Natalie A. Lissy,et al.  Killing HIV-infected cells by transduction with an HIV protease-activated caspase-3 protein , 1999, Nature Medicine.