tailieunhanh - Báo cáo khoa học: Different mechanisms for cellular internalization of the HIV-1 Tat-derived cell penetrating peptide and recombinant proteins fused to Tat

Translocation through the plasma membrane is a major limiting step for the cellular delivery of macromolecules. A promising strategy to overcome this problem consists in the chemical conjugation (or fusion) to cell penetrating peptides (CPP) derived from proteins able to cross the plasma membrane. A large number of di erent cargo mol-ecules such as oligonucleotides, peptides, peptide nucleic acids, proteins or even nanoparticles have been internalized in cells by this strategy. | Eur. J. Biochem. 269 494-501 2002 FEBS 2002 Different mechanisms for cellular internalization of the HIV-1 Tat-derived cell penetrating peptide and recombinant proteins fused to Tat Michelle Silhol1 Mudit Tyagi2 Mauro Giacca2 Bernard Lebleu1 and Eric Vives1 1Institut de Génétique Moleculaire de Montpellier CNRS UMR 5124 BP5051 Montpellier France 2Molecular Medicine Laboratory International Centre for Genetic Engineering and Biotechnology Trieste Italy Translocation through the plasma membrane is a major limiting step for the cellular delivery of macromolecules. A promising strategy to overcome this problem consists in the chemical conjugation or fusion to cell penetrating peptides CPP derived from proteins able to cross the plasma membrane. A large number of different cargo molecules such as oligonucleotides peptides peptide nucleic acids proteins or even nanoparticles have been internalized in cells by this strategy. One of these translocating peptides was derived from the HIV-1 Tat protein. The mechanisms by which CPP enter cells remain unknown. Recently convincing biochemical and genetic findings has established that the full-length Tat protein was internalized in cells via the ubiquitous heparan sulfate HS proteoglycans. We demonstrate here that the short Tat CPP is taken up by a route that does not involve the HS proteoglycans. Keywords Tat cell penetrating peptide CPP cellular uptake heparan sulfate. Several cell-penetrating peptides CPP allowing the efficient internalization of various nonpermeant drugs in different cell lines have been recently described. A covalent link had to be created between the CPP and the cargo molecule to promote efficient membrane translocation of the chimera 1-7 . A 16-mer peptide derived from the Antennapedia protein homeodomain 8 and a 13-mer peptide derived from the HIV-1 Tat protein 9 have been extensively studied. In our initial experiments using the short Tat basic domain we demonstrated the uptake of chemically conjugated .