Cellular uptake of S413-PV peptide occurs upon conformational changes induced by peptide-membrane interactions

Miguel Mano; Ana Henriques; Artur Paiva; Manuel Prieto; Francisco Gavilanes; Sérgio Simões; Maria C. Pedroso de Lima

doi:10.1016/j.bbamem.2006.01.014

Cellular uptake of S4₁₃-PV peptide occurs upon conformational changes induced by peptide-membrane interactions

Miguel Mano, Ana Henriques, Artur Paiva, Manuel Prieto, Francisco Gavilanes, Sérgio Simões, Maria C. Pedroso de Lima^*

^*Corresponding author for this work

Cardiovascular Sciences

Research output: Contribution to journal › Article › peer-review

29 Citations (Scopus)

Abstract

In face of accumulated reports demonstrating that uptake of some cell-penetrating peptides occurs through previously described endocytic pathways, or is a consequence of cell fixation artifacts, we conducted a systematic analysis on the mechanism responsible for the cellular uptake of the S4₁₃-PV karyophilic cell-penetrating peptide. The results reviewed here show that the S4₁₃-PV peptide is able to very efficiently accumulate inside live cells in a rapid, non-toxic and dose-dependent manner, through a mechanism distinct from endocytosis. Comparative analysis of peptide uptake by mutant cells lacking heparan sulfate proteoglycans demonstrates that, although not mandatory, their presence at cell surface facilitates the cellular uptake of the S4₁₃-PV peptide. Furthermore, we demonstrate that upon interaction with lipid vesicles, the S4₁₃-PV peptide undergoes significant conformational changes that are consistent with the formation of helical structures. Such conformational changes occur concomitantly with a penetration of the peptide into the lipid bilayer, strongly suggesting that the resulting helical structures are crucial for the non-endocytic cellular uptake of the S4₁₃-PV peptide. Overall, our data support that, rather than endocytosis, the cellular uptake of the S4₁₃-PV cell-penetrating peptide is a consequence of its direct translocation through cell membranes following conformational changes induced by peptide-membrane interactions.

Original language	English
Pages (from-to)	336-346
Number of pages	11
Journal	Biochimica et Biophysica Acta - Biomembranes
Volume	1758
Issue number	3
DOIs	https://doi.org/10.1016/j.bbamem.2006.01.014
Publication status	Published - Mar 2006

Keywords

Amphipathic alpha-helix
Cell-penetrating peptide
Circular dichroism
Peptide-membrane interaction
Protein transduction domain
Tryptophan fluorescence

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10.1016/j.bbamem.2006.01.014

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title = "Cellular uptake of S413-PV peptide occurs upon conformational changes induced by peptide-membrane interactions",

abstract = "In face of accumulated reports demonstrating that uptake of some cell-penetrating peptides occurs through previously described endocytic pathways, or is a consequence of cell fixation artifacts, we conducted a systematic analysis on the mechanism responsible for the cellular uptake of the S413-PV karyophilic cell-penetrating peptide. The results reviewed here show that the S413-PV peptide is able to very efficiently accumulate inside live cells in a rapid, non-toxic and dose-dependent manner, through a mechanism distinct from endocytosis. Comparative analysis of peptide uptake by mutant cells lacking heparan sulfate proteoglycans demonstrates that, although not mandatory, their presence at cell surface facilitates the cellular uptake of the S413-PV peptide. Furthermore, we demonstrate that upon interaction with lipid vesicles, the S413-PV peptide undergoes significant conformational changes that are consistent with the formation of helical structures. Such conformational changes occur concomitantly with a penetration of the peptide into the lipid bilayer, strongly suggesting that the resulting helical structures are crucial for the non-endocytic cellular uptake of the S413-PV peptide. Overall, our data support that, rather than endocytosis, the cellular uptake of the S413-PV cell-penetrating peptide is a consequence of its direct translocation through cell membranes following conformational changes induced by peptide-membrane interactions.",

keywords = "Amphipathic alpha-helix, Cell-penetrating peptide, Circular dichroism, Peptide-membrane interaction, Protein transduction domain, Tryptophan fluorescence",

author = "Miguel Mano and Ana Henriques and Artur Paiva and Manuel Prieto and Francisco Gavilanes and S{\'e}rgio Sim{\~o}es and {Pedroso de Lima}, {Maria C.}",

note = "Funding Information: We thank Prof. F. Regateiro, Head of the Centro de Histocompatibilidade de Coimbra (Portugal), for scientific collaboration in this study. This study was supported by a grant from the Portuguese Foundation for Science and Technology (POCTI/CVT/44854/2002). M. Mano is recipient of a fellowship from the Portuguese Foundation for Science and Technology.",

year = "2006",

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volume = "1758",

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T1 - Cellular uptake of S413-PV peptide occurs upon conformational changes induced by peptide-membrane interactions

AU - Mano, Miguel

AU - Henriques, Ana

AU - Paiva, Artur

AU - Prieto, Manuel

AU - Gavilanes, Francisco

AU - Simões, Sérgio

AU - Pedroso de Lima, Maria C.

N1 - Funding Information: We thank Prof. F. Regateiro, Head of the Centro de Histocompatibilidade de Coimbra (Portugal), for scientific collaboration in this study. This study was supported by a grant from the Portuguese Foundation for Science and Technology (POCTI/CVT/44854/2002). M. Mano is recipient of a fellowship from the Portuguese Foundation for Science and Technology.

PY - 2006/3

Y1 - 2006/3

N2 - In face of accumulated reports demonstrating that uptake of some cell-penetrating peptides occurs through previously described endocytic pathways, or is a consequence of cell fixation artifacts, we conducted a systematic analysis on the mechanism responsible for the cellular uptake of the S413-PV karyophilic cell-penetrating peptide. The results reviewed here show that the S413-PV peptide is able to very efficiently accumulate inside live cells in a rapid, non-toxic and dose-dependent manner, through a mechanism distinct from endocytosis. Comparative analysis of peptide uptake by mutant cells lacking heparan sulfate proteoglycans demonstrates that, although not mandatory, their presence at cell surface facilitates the cellular uptake of the S413-PV peptide. Furthermore, we demonstrate that upon interaction with lipid vesicles, the S413-PV peptide undergoes significant conformational changes that are consistent with the formation of helical structures. Such conformational changes occur concomitantly with a penetration of the peptide into the lipid bilayer, strongly suggesting that the resulting helical structures are crucial for the non-endocytic cellular uptake of the S413-PV peptide. Overall, our data support that, rather than endocytosis, the cellular uptake of the S413-PV cell-penetrating peptide is a consequence of its direct translocation through cell membranes following conformational changes induced by peptide-membrane interactions.

AB - In face of accumulated reports demonstrating that uptake of some cell-penetrating peptides occurs through previously described endocytic pathways, or is a consequence of cell fixation artifacts, we conducted a systematic analysis on the mechanism responsible for the cellular uptake of the S413-PV karyophilic cell-penetrating peptide. The results reviewed here show that the S413-PV peptide is able to very efficiently accumulate inside live cells in a rapid, non-toxic and dose-dependent manner, through a mechanism distinct from endocytosis. Comparative analysis of peptide uptake by mutant cells lacking heparan sulfate proteoglycans demonstrates that, although not mandatory, their presence at cell surface facilitates the cellular uptake of the S413-PV peptide. Furthermore, we demonstrate that upon interaction with lipid vesicles, the S413-PV peptide undergoes significant conformational changes that are consistent with the formation of helical structures. Such conformational changes occur concomitantly with a penetration of the peptide into the lipid bilayer, strongly suggesting that the resulting helical structures are crucial for the non-endocytic cellular uptake of the S413-PV peptide. Overall, our data support that, rather than endocytosis, the cellular uptake of the S413-PV cell-penetrating peptide is a consequence of its direct translocation through cell membranes following conformational changes induced by peptide-membrane interactions.

KW - Amphipathic alpha-helix

KW - Cell-penetrating peptide

KW - Circular dichroism

KW - Peptide-membrane interaction

KW - Protein transduction domain

KW - Tryptophan fluorescence

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AN - SCOPUS:33646534241

SN - 0005-2736

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EP - 346

JO - Biochimica et Biophysica Acta - Biomembranes

JF - Biochimica et Biophysica Acta - Biomembranes

IS - 3

ER -

Cellular uptake of S4₁₃-PV peptide occurs upon conformational changes induced by peptide-membrane interactions

Abstract

Keywords

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