Effects of Tryptophan Content and Backbone Spacing on the Uptake Efficiency of Cell-Penetrating Peptides
Artikel i vetenskaplig tidskrift, 2012

Cell-penetrating peptides (CPPs) are able to traverse cellular membranes and deliver macromolecular cargo. Uptake occurs through both endocytotic and nonendocytotic pathways, but the molecular requirements for efficient internalization are not fully understood. Here we investigate how the presence of tryptophans and their position within an oligoarginine influence uptake mechanism and efficiency. Flow cytometry and confocal fluorescence imaging are used to estimate uptake efficiency, intracellular distribution and toxicity in Chinese hamster ovarian cells. Further, membrane leakage and lipid membrane affinity are investigated. The peptides contain eight arginine residues and one to four tryptophans, the tryptophans positioned either at the N-terminus, in the middle, or evenly distributed along the amino acid sequence. Our data show that the intracellular distribution varies among peptides with different tryptophan content and backbone spacing. Uptake efficiency is higher for the peptides with four tryptophans in the middle, or evenly distributed along the peptide sequence, than for the peptide with four tryptophans at the N-terminus. All peptides display low cytotoxicity except for the one with four tryptophans at the N-terminus, which was moderately toxic. This finding is consistent with their inability to induce efficient leakage of dye from lipid vesicles. All peptides have comparable affinities for lipid vesicles, showing that lipid binding is not a decisive parameter for uptake. Our results indicate that tryptophan content and backbone spacing can affect both the CPP uptake efficiency and the CPP uptake mechanism. The low cytotoxicity of these peptides and the possibilities of tuning their uptake mechanism are interesting from a therapeutic point of view.

arginine-rich peptides

delivery

mechanisms

endocytosis

transporters

plasma-membrane

binding

internalization

translocation

membrane-proteins

molecular

Författare

Hanna Rydberg

Chalmers, Kemi- och bioteknik, Fysikalisk kemi

Maria Matson Dzebo

Chalmers, Kemi- och bioteknik, Fysikalisk kemi

Helene Åmand

Chalmers, Kemi- och bioteknik, Fysikalisk kemi

Elin Esbjörner Winters

Chalmers, Kemi- och bioteknik, Fysikalisk kemi

Bengt Nordén

Chalmers, Kemi- och bioteknik, Fysikalisk kemi

Biochemistry

0006-2960 (ISSN) 1520-4995 (eISSN)

Vol. 51 5531-5539

Styrkeområden

Energi

Livsvetenskaper och teknik

Materialvetenskap

Ämneskategorier

Kemi

DOI

10.1021/bi300454k