Effects of Tryptophan Content and Backbone Spacing on the Uptake Efficiency of Cell-Penetrating Peptides

Handle URI:
http://hdl.handle.net/10754/598092
Title:
Effects of Tryptophan Content and Backbone Spacing on the Uptake Efficiency of Cell-Penetrating Peptides
Authors:
Rydberg, Hanna A.; Matson, Maria; Åmand, Helene L.; Esbjörner, Elin K.; Nordén, Bengt
Abstract:
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. © 2012 American Chemical Society.
Citation:
Rydberg HA, Matson M, Åmand HL, Esbjörner EK, Nordén B (2012) Effects of Tryptophan Content and Backbone Spacing on the Uptake Efficiency of Cell-Penetrating Peptides. Biochemistry 51: 5531–5539. Available: http://dx.doi.org/10.1021/bi300454k.
Publisher:
American Chemical Society (ACS)
Journal:
Biochemistry
Issue Date:
10-Jul-2012
DOI:
10.1021/bi300454k
PubMed ID:
22712882
Type:
Article
ISSN:
0006-2960; 1520-4995
Sponsors:
This work was supported by a grant to B.N. from King Abdullah University of Science and Technology (KAUST) and to E.K.E. from the Lennander foundation.
Appears in Collections:
Publications Acknowledging KAUST Support

Full metadata record

DC FieldValue Language
dc.contributor.authorRydberg, Hanna A.en
dc.contributor.authorMatson, Mariaen
dc.contributor.authorÅmand, Helene L.en
dc.contributor.authorEsbjörner, Elin K.en
dc.contributor.authorNordén, Bengten
dc.date.accessioned2016-02-25T13:12:31Zen
dc.date.available2016-02-25T13:12:31Zen
dc.date.issued2012-07-10en
dc.identifier.citationRydberg HA, Matson M, Åmand HL, Esbjörner EK, Nordén B (2012) Effects of Tryptophan Content and Backbone Spacing on the Uptake Efficiency of Cell-Penetrating Peptides. Biochemistry 51: 5531–5539. Available: http://dx.doi.org/10.1021/bi300454k.en
dc.identifier.issn0006-2960en
dc.identifier.issn1520-4995en
dc.identifier.pmid22712882en
dc.identifier.doi10.1021/bi300454ken
dc.identifier.urihttp://hdl.handle.net/10754/598092en
dc.description.abstractCell-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. © 2012 American Chemical Society.en
dc.description.sponsorshipThis work was supported by a grant to B.N. from King Abdullah University of Science and Technology (KAUST) and to E.K.E. from the Lennander foundation.en
dc.publisherAmerican Chemical Society (ACS)en
dc.titleEffects of Tryptophan Content and Backbone Spacing on the Uptake Efficiency of Cell-Penetrating Peptidesen
dc.typeArticleen
dc.identifier.journalBiochemistryen
dc.contributor.institutionChalmers University of Technology, Göteborg, Swedenen

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