Development of a Cell-penetrating Peptide that Exhibits Responsive Changes in its Secondary Structure in the Cellular Environment.
Cell-penetrating peptides (CPP) are received a lot of attention as an intracellular delivery tool for hydrophilic molecules such as drugs, proteins, and DNAs. We designed and synthesized nona-arginine analogues 1-5 [FAM-β-Ala-(l-Arg-l-Arg-l-Pro)3-(Gly)3-NH2 (1), FAM-β-Ala-(l-Arg-l-Arg-l-Pro(NH2))3-(Gly)3-NH2 (2), FAM-β-Ala-(l-Arg-l-Arg-l-Pro(Gu))3-(Gly)3-NH2 (3), FAM-β-Ala-(l-Arg)2-(l-Pro(Gu))2-(l-Arg)4-l-Pro(Gu)-(Gly)3-NH2 (4), and FAM-β-Ala-(l-Arg)6-(l-Pro(Gu))3-(Gly)3-NH2 (5)] containing l-proline (l-Pro) or cationic proline derivatives (l-Pro(NH2) and l-Pro(Gu)), and investigated their cell-penetrating abilities. Interestingly, only peptide 3 having the side-chain guanidinyl l-Pro(Gu) exhibited a secondary structural change in cellular environment. Specifically, peptide 3 formed a random structure in hydrophilic conditions, whereas it formed a helical structure under amphipathic conditions. Furthermore, during cellular permeability tests, peptide 3 demonstrated greater cell-penetrating activity than other peptides and effectively transported plasmid DNA into HeLa cells. Thus, l-Pro(Gu)-containing peptide 3 may be a useful candidate as a gene delivery carrier.
National Institute of Health Sciences, Tokyo 158-8501, Japan.
Hiroko
H
Yamashita
Graduate School of Biomedical Sciences, Nagasaki University, 1-14 Bunkyo-machi, Nagasaki 852-8521, Japan.
Takuma
T
Kato
Graduate School of Biomedical Sciences, Nagasaki University, 1-14 Bunkyo-machi, Nagasaki 852-8521, Japan.
Makoto
M
Oba
National Institute of Health Sciences, Tokyo 158-8501, Japan.
Takashi
T
Misawa
National Institute of Health Sciences, Tokyo 158-8501, Japan.
Takayuki
T
Hattori
National Institute of Health Sciences, Tokyo 158-8501, Japan.
Nobumichi
N
Ohoka
Graduate School of Biomedical Sciences, Nagasaki University, 1-14 Bunkyo-machi, Nagasaki 852-8521, Japan.
Masakazu
M
Tanaka
National Institute of Health Sciences, Tokyo 158-8501, Japan.
Mikihiko
M
Naito
National Institute of Health Sciences, Tokyo 158-8501, Japan.
Masaaki
M
Kurihara
National Institute of Health Sciences, Tokyo 158-8501, Japan.
Yosuke
Y
Demizu
D057846:Cell-Penetrating Peptides / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism*; D004247:DNA / Q000378:metabolism; D004705:Endocytosis; D006367:HeLa Cells; D006801:Humans; D010957:Plasmids / Q000378:metabolism; D017433:Protein Structure, Secondary
D057846:Cell-Penetrating Peptides; D004247:DNA