Dual peptide conjugation strategy for improved cellular uptake and mitochondria targeting

Ran Lin, Pengcheng Zhang, Andrew G. Cheetham, Jeremy D Walston, Peter Abadir, Honggang Cui

Research output: Contribution to journalArticle

Abstract

Mitochondria are critical regulators of cellular function and survival. Delivery of therapeutic and diagnostic agents into mitochondria is a challenging task in modern pharmacology because the molecule to be delivered needs to first overcome the cell membrane barrier and then be able to actively target the intracellular organelle. Current strategy of conjugating either a cell penetrating peptide (CPP) or a subcellular targeting sequence to the molecule of interest only has limited success. We report here a dual peptide conjugation strategy to achieve effective delivery of a non-membrane-penetrating dye 5-carboxyfluorescein (5-FAM) into mitochondria through the incorporation of both a mitochondrial targeting sequence (MTS) and a CPP into one conjugated molecule. Notably, circular dichroism studies reveal that the combined use of α-helix and PPII-like secondary structures has an unexpected, synergistic contribution to the internalization of the conjugate. Our results suggest that although the use of positively charged MTS peptide allows for improved targeting of mitochondria, with MTS alone it showed poor cellular uptake. With further covalent linkage of the MTS-5-FAM conjugate to a CPP sequence (R8), the dually conjugated molecule was found to show both improved cellular uptake and effective mitochondria targeting. We believe these results offer important insight into the rational design of peptide conjugates for intracellular delivery. (Chemical Equation Presented).

Original languageEnglish (US)
Pages (from-to)71-77
Number of pages7
JournalBioconjugate Chemistry
Volume26
Issue number1
DOIs
StatePublished - Jan 21 2015

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Mitochondria
Peptides
Cell-Penetrating Peptides
Molecules
Cell membranes
Circular Dichroism
Organelles
Dichroism
Coloring Agents
Cell Membrane
Pharmacology
Dyes

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Organic Chemistry
  • Pharmaceutical Science
  • Biomedical Engineering
  • Pharmacology

Cite this

Dual peptide conjugation strategy for improved cellular uptake and mitochondria targeting. / Lin, Ran; Zhang, Pengcheng; Cheetham, Andrew G.; Walston, Jeremy D; Abadir, Peter; Cui, Honggang.

In: Bioconjugate Chemistry, Vol. 26, No. 1, 21.01.2015, p. 71-77.

Research output: Contribution to journalArticle

Lin, Ran ; Zhang, Pengcheng ; Cheetham, Andrew G. ; Walston, Jeremy D ; Abadir, Peter ; Cui, Honggang. / Dual peptide conjugation strategy for improved cellular uptake and mitochondria targeting. In: Bioconjugate Chemistry. 2015 ; Vol. 26, No. 1. pp. 71-77.
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