Cellular internalization of arginine-rich peptides into tobacco suspension cells: a structure-activity relationship study.
Translocation of several fluorescently labeled arginine-rich peptides into intact plant cells was quantitatively examined in order to investigate the structural factors required for efficient cellular internalization, and thereby, to evaluate the potential of arginine-rich peptides as intracellular delivery vectors in plants. Cell-penetrating peptides (CPPs) such as arginine-rich peptides permit the direct introduction of biologically active macromolecules into plant cytoplasm to manipulate various intracellular processes. While a significant level of adsorption of applied arginine-rich peptides was observed in the cell walls rich in negative charges, removal of adsorbed peptides by trypsin treatment allowed determination of the amount of internalized peptides in a quantitative manner using spectrofluorometric analysis. The internalization of arginine-rich peptides depended on the number of arginine residues, and the peptide containing eight arginine residues showed most effective internalization. Besides, the position of small cargoes attached to the arginine-rich peptides markedly affected the internalization efficiency. The results obtained in this study provide useful information for the development of efficient intracellular delivery tools in plant science.
Mizuno
Takashi
T
Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kyoto 606-8502, Japan.
Miyashita
Masahiro
M
Miyagawa
Hisashi
H
D001120:Arginine / Q000737:chemistry; D001693:Biological Transport, Active; D002473:Cell Wall / Q000378:metabolism; D002478:Cells, Cultured; D016503:Drug Delivery Systems; D005452:Fluoresceins; D005456:Fluorescent Dyes; D008856:Microscopy, Fluorescence; D009842:Oligopeptides / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism*; D013050:Spectrometry, Fluorescence; D013329:Structure-Activity Relationship; D014026:Nicotiana / Q000378:metabolism*
D005452:Fluoresceins; D005456:Fluorescent Dyes; D009842:Oligopeptides; D001120:Arginine; C018506:diacetylfluorescein