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|Título:||Kinetic uptake profiles of cell penetrating peptides in lymphocytes and monocytes|
de la Torre, Beatriz G.
Santos, Nuno C.
|Palavras-chave:||Nucleolar targeting peptide|
Cell penetrating peptide
|Citação:||Biochimica et Biophysica Acta 1830 (2013) 4554–4563|
|Resumo:||Background: Nucleolar targeting peptides (NrTPs), resulting from structural minimization of the rattlesnake toxin crotamine, are a novel family of cell-penetrating peptides (CPPs) shown to internalize and deliver cargos into different cell types. Methods: In this study, we address NrTP kinetics of translocation into primary cells. We used flow cytometry to measure the intracellular uptake of rhodamine B-labeled NrTPs in peripheral blood mononucleated cells (PBMCs). Results: The kinetic profiles for each peptide are concentration-independent but significantly different among NrTPs, pointing out for the amino acid sequence importance. Arginine-containing peptides (NrTP7 and Tat48–60, used for comparison) were found to be more toxic than lysine-containing ones, as expected. On the other hand, one same peptide behaves differently in each of the lymphocyte and monocyte cell populations, suggesting differences in entry mechanism that in turn reflect diversity in cell functionality. Uptake results obtained at 4 °C or using chemical endocytosis inhibitors support the importance of non-endocytic mechanisms in the cellular internalization of NrTP1 and NrTP5, while confirming endocytosis as the main mechanism of NrTPs entry. Conclusion: Overall, both direct translocation and endocytosis mechanisms play a role in NrTP entry. Yet, there is predominance of endocytosis-mediated mechanisms. NrTPs (especially NrTP6) are an excellent intracellular delivery tool, with efficient internalization and no toxicity. General significance: This work validates NrTPs as potential therapeutic tools for, e.g., cancer or inhibition of viral replication and establishes a new comparative and quantitative method to test CPP efficiency.|
|Descrição:||© 2013 Elsevier B.V. All rights reserved.|
|Versão do Editor:||The definitive version is available at http://www.elsevier.com|
|Aparece nas colecções:||IMM - Artigos em Revistas Internacionais|
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