Quantitative comparison of membrane transduction and endocytosis of oligopeptides

A novel method was developed to quantitatively determine the membrane transduction efficiency of cationic oligopeptides, allowing for selective measurement of transduction, excluding concurrent adsorptive endocytosis. It was found that Tat-(47–57) (YGRKKRRQRRR), YG(l-R) 9, and guanidinated-YG(l-K) 9...

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Bibliographic Details
Published in:Biochemical and biophysical research communications 2003-07, Vol.307 (2), p.241-247
Main Authors: Zaro, Jennica L., Shen, Wei-Chiang
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Animals
Arginine - chemistry
Arginine - metabolism
Biological Transport - physiology
Cationic oligopeptides
Cell Fractionation - methods
CHO Cells
Cricetinae
Culture Media, Serum-Free
Endocytosis - physiology
Gene Products, tat - genetics
Gene Products, tat - metabolism
Iodine Radioisotopes - metabolism
Lysine - chemistry
Lysine - metabolism
Membrane transduction
Oligopeptides - genetics
Oligopeptides - metabolism
Peptide Fragments - genetics
Peptide Fragments - metabolism
Quantitative measurement
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Summary:A novel method was developed to quantitatively determine the membrane transduction efficiency of cationic oligopeptides, allowing for selective measurement of transduction, excluding concurrent adsorptive endocytosis. It was found that Tat-(47–57) (YGRKKRRQRRR), YG(l-R) 9, and guanidinated-YG(l-K) 9 were preferentially transduced to the cytosolic compartment, while YG(l-K) 9 was primarily endocytosed. Studies of various oligoarginine peptides (4–15 residues) demonstrated that internalization through transduction remained constant, while the amount internalized via endocytosis increased with arginine length, indicating that oligopeptide transduction requires the guanidine structure of arginine, while endocytosis depends only on the number of positive charges. The addition of unlabeled-YG(l-R) 9 dose-dependently inhibited transduction of 125I-labeled-YG(l-R) 9, while endocytosis remained constant. Unlabeled-YG(d-R) 9 and Tat-(47–57) were able to inhibit transduction similarly, while YG(l-K) 9 had no effect. These studies demonstrate specific surface binding sites required for transduction of oligoarginine, but not for endocytosis. Data generated from this study are essential to elucidate the transduction mechanism and to develop an efficient carrier system for cytoplasmic drug delivery.
ISSN:0006-291X
1090-2104
DOI:10.1016/S0006-291X(03)01167-7