Cellular uptake of Antennapedia Penetratin peptides is a two-step process in which phase transfer precedes a tryptophan-dependent translocation
| Autor: | Olivier Bouffioux, Alain Prochiantz, Jacques J. Picard, Chloë Shaw-Jackson, Robert Brasseur, Christelle Matis, Marie-Paule Mingeot-Leclercq, Geneviève Dom, René Rezsohazy |
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| Přispěvatelé: | UCL - MD/FARM - Ecole de pharmacie, UCL - MD/MIGE - Département de microbiologie, d'immunologie et de génétique |
| Jazyk: | angličtina |
| Rok vydání: | 2003 |
| Předmět: |
media_common.quotation_subject
Hydrophobicity Oligonucleotides Peptide Cell-Penetrating Peptides Biology Antennapedia Genetics Animals Amino Acid Sequence Internalization Peptide sequence media_common chemistry.chemical_classification COS cells Tryptophan Temperature Water Biological membrane Articles Membrane Biochemistry chemistry Amino Acid Substitution COS Cells Mutation Biophysics Chloroform Carrier Proteins Hydrophobic and Hydrophilic Interactions |
| Zdroj: | Nucleic Acids Research, Vol. 31, no. 2, p. 556-561 (2003) |
| Popis: | Several homeodomains and homeodomain-containing proteins enter live cells through a receptor- and energy-independent mechanism. Translocation through biological membranes is conferred by the third alpha-helix of the homeodomain, also known as Penetratin. Biophysical studies demonstrate that entry of Penetratin into cells requires its binding to surface lipids but that binding and translocation are differentially affected by modifications of some physico-chemical properties of the peptide, like helical amphipathicity or net charge. This suggests that the plasma membrane lipid composition affects the internalization of Penetratin and that internalization requires both lipid binding and other specific properties. Using a phase transfer assay, it is shown that negatively charged lipids promote the transfer of Penetratin from a hydrophilic into a hydrophobic environment, probably through charge neutralization. Accordingly, transfer into a hydrophobic milieu can also be obtained in the absence of negatively charged lipids, by the addition of DNA oligonucleotides. Strikingly, phase transfer by charge neutralization was also observed with a variant peptide of same charge and hydrophobicity in which the tryptophan at position 6 was replaced by a phenylalanine. However, Penetratin, but not its mutant version, is internalized by live cells. This underscores that charge neutralization and phase transfer represent only a first step in the internalization process and that further crossing of a biological membrane necessitates the critical tryptophan residue at position 6. |
| Databáze: | OpenAIRE |
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