On the cellular uptake and membrane effect of the multifunctional peptide, TatLK15

Myasar Alkotaji, Alain Pluen, Egor Zindy, Zahra Hamrang, Harmesh Aojula

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Recently, the multifunctional peptide TatLK15 resulting from the fusion of the cell penetrating peptide Tat and the amphipathic peptide LK15 was shown to be efficient at mediating siRNA and shRNA delivery in leukemia cells to silence the bcr-abl oncoprotein. The present study focused on TatLK15 peptide cellular uptake and defining conditions for its use within a range of doses exhibiting minimal toxicity. The initial part of the study carried out in solution confirmed that the insertion of a glycine bridge allowed retention of the LK15 α-helicity, and fluorescence correlation spectroscopy did not reveal preferential conformations at the studied concentrations. In the second part, TatLK15 uptake mechanisms appeared peptide dose- and cell line- dependent as well as requiring membrane potential. Below a critical dose, TatLK15 toxicity appeared limited for approximately three hours as demonstrated by the combined use of lactate dehydrogenase release, MTT assays, and time-dependent observation of membrane-impermeant dye uptake using high content screening apparatus. Furthermore, toxicity was observed to occur rapidly at higher peptide doses. Finally, a comparison between TatLK15 and another Tat amphipathic peptide construct suggested that α-helix content should be viewed as a key element in the development of similar peptides.
Original languageEnglish
Pages (from-to)293-304
Number of pages12
JournalJournal of Pharmaceutical Sciences
Volume103
Issue number1
DOIs
Publication statusPublished - 1 Jan 2014

Fingerprint

Peptides
Membranes
Small Interfering RNA
Cell-Penetrating Peptides
Oncogene Proteins
Fluorescence Spectrometry
L-Lactate Dehydrogenase
Membrane Potentials
Glycine
Leukemia
Coloring Agents
Observation
Cell Line

Keywords

  • cell penetrating peptide
  • peptide delivery
  • Tat
  • membrane translocation
  • diffusion
  • toxicity
  • image analysis
  • fluorescence spectroscopy
  • circular dichroism

Cite this

Alkotaji, Myasar ; Pluen, Alain ; Zindy, Egor ; Hamrang, Zahra ; Aojula, Harmesh. / On the cellular uptake and membrane effect of the multifunctional peptide, TatLK15. In: Journal of Pharmaceutical Sciences. 2014 ; Vol. 103, No. 1. pp. 293-304.
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abstract = "Recently, the multifunctional peptide TatLK15 resulting from the fusion of the cell penetrating peptide Tat and the amphipathic peptide LK15 was shown to be efficient at mediating siRNA and shRNA delivery in leukemia cells to silence the bcr-abl oncoprotein. The present study focused on TatLK15 peptide cellular uptake and defining conditions for its use within a range of doses exhibiting minimal toxicity. The initial part of the study carried out in solution confirmed that the insertion of a glycine bridge allowed retention of the LK15 α-helicity, and fluorescence correlation spectroscopy did not reveal preferential conformations at the studied concentrations. In the second part, TatLK15 uptake mechanisms appeared peptide dose- and cell line- dependent as well as requiring membrane potential. Below a critical dose, TatLK15 toxicity appeared limited for approximately three hours as demonstrated by the combined use of lactate dehydrogenase release, MTT assays, and time-dependent observation of membrane-impermeant dye uptake using high content screening apparatus. Furthermore, toxicity was observed to occur rapidly at higher peptide doses. Finally, a comparison between TatLK15 and another Tat amphipathic peptide construct suggested that α-helix content should be viewed as a key element in the development of similar peptides.",
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On the cellular uptake and membrane effect of the multifunctional peptide, TatLK15. / Alkotaji, Myasar; Pluen, Alain; Zindy, Egor; Hamrang, Zahra; Aojula, Harmesh.

In: Journal of Pharmaceutical Sciences, Vol. 103, No. 1, 01.01.2014, p. 293-304.

Research output: Contribution to journalArticle

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