Synthesis and studies on cell-penetrating peptides.
The ability of different synthetic cell penetrating peptides, as Antennapedia (wild and Phe(6) mutated penetratins), flock house virus, and integrin peptides to form complexes with a 25mer antisense oligonucleotide was compared and their conformation was determined by circular dichroism spectroscopy. The efficiency for oligonucleotide delivery into cells was measured using peptides labeled with a coumarin derivative showing blue fluorescence and the fluorescein-labeled antisense oligonucleotide showing green fluorescence. Fluorescence due to the excitation energy transfer confirmed the interaction of the antisense oligonucleotide and cell-penetrating peptides. The most efficient oligonucleotide delivery was found for penetratins. Comparison of the two types of penetratins shows that the wild-type penetratin proved to be more efficient than mutated penetratin. The paper also emphasizes that the attachment of a fluorescent label may have an effect on the conformation and flexibility of cell-penetrating peptides that must be taken into consideration when evaluating biological experiments.
Bertrand
Jean-Remi
JR
Institute Gustave Roussy, CNRS UMR 8121, University Paris 11, Villejuif, France.
Malvy
Claude
C
Auguste
Tiphanie
T
Tóth
Gábor K
GK
Kiss-Ivánkovits
Orsolya
O
Illyés
Eszter
E
Hollósi
Miklós
M
Bottka
Sándor
S
Laczkó
Ilona
I
D000595:Amino Acid Sequence; D000818:Animals; D002352:Carrier Proteins / Q000138:chemical synthesis / Q000737:chemistry* / Q000235:genetics / Q000493:pharmacokinetics*; D002461:Cell Line, Transformed; D057846:Cell-Penetrating Peptides; D002942:Circular Dichroism; D015536:Down-Regulation; D004337:Drug Carriers / Q000138:chemical synthesis / Q000737:chemistry* / Q000493:pharmacokinetics*; D018014:Gene Transfer Techniques; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation; D041681:NIH 3T3 Cells; D016376:Oligonucleotides, Antisense / Q000737:chemistry / Q000235:genetics / Q000493:pharmacokinetics*; D015514:Oncogene Proteins, Fusion / Q000235:genetics*; D011487:Protein Conformation; D021381:Protein Transport; D051789:Proto-Oncogene Protein c-fli-1 / Q000235:genetics*; D034802:RNA-Binding Protein EWS
D002352:Carrier Proteins; D057846:Cell-Penetrating Peptides; D004337:Drug Carriers; C106031:EWS-FLI fusion protein; D016376:Oligonucleotides, Antisense; D015514:Oncogene Proteins, Fusion; D051789:Proto-Oncogene Protein c-fli-1; D034802:RNA-Binding Protein EWS; C414409:penetratin