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Simultaneous membrane interaction of amphipathic peptide monomers, self-aggregates and cargo complexes detected by fluorescence correlation spectroscopy
Stockholm University, Faculty of Science, Department of Neurochemistry.ORCID iD: 0000-0001-7522-8964
Stockholm University, Faculty of Science, Department of Neurochemistry.
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Number of Authors: 7
2018 (English)In: Biochimica et Biophysica Acta - Biomembranes, ISSN 0005-2736, E-ISSN 1879-2642, Vol. 1860, no 2, p. 491-504Article in journal (Refereed) Published
Abstract [en]

Peptides able to translocate cell membranes while carrying macromolecular cargo, as cell-penetrating peptides (CPPs), can contribute to the field of drug delivery by enabling the transport of otherwise membrane impermeable molecules. Formation of non-covalent complexes between amphipathic peptides and oligonucleotides is driven by electrostatic and hydrophobic interactions. Here we investigate and quantify the coexistence of distinct molecular species in multiple equilibria, namely peptide monomer, peptide self-aggregates and peptide/oligonucleotide complexes. As a model for the complexes, we used a stearylated peptide from the PepFect family, PF14 and siRNA. PF14 has a cationic part and a lipid part, resembling some characteristics of cationic lipids. Fluorescence correlation spectroscopy (FCS) and fluorescence cross-correlation spectroscopy (FCCS) were used to detect distinct molecular entities in solution and at the plasma membrane of live cells. For that, we labeled the peptide with carboxyrhodamine 6G and the siRNA with Cyanine 5. We were able to detect fluorescent entities with diffusional properties characteristic of the peptide monomer as well as of peptide aggregates and peptide/oligonucleotide complexes. Strategies to avoid peptide adsorption to solid surfaces and self-aggregation were developed and allowed successful FCS measurements in solution and at the plasma membrane. The ratio between the detected molecular species was found to vary with pH, peptide concentration and the proximity to the plasma membrane. The present results suggest that the diverse cellular uptake mechanisms, often reported for amphipathic CPPs, might result from the synergistic effect of peptide monomers, self-aggregates and cargo complexes, distributed unevenly at the plasma membrane.

Place, publisher, year, edition, pages
2018. Vol. 1860, no 2, p. 491-504
Keyword [en]
Cell-penetrating peptide, Fluorescence correlation spectroscopy, PepFect14, siRNA, Amphipathic peptide, Peptide aggregates, Plasma membrane
National Category
Biological Sciences
Identifiers
URN: urn:nbn:se:su:diva-153640DOI: 10.1016/j.bbamem.2017.09.024ISI: 000424183500029PubMedID: 28962904OAI: oai:DiVA.org:su-153640DiVA, id: diva2:1188852
Available from: 2018-03-08 Created: 2018-03-08 Last updated: 2018-03-08Bibliographically approved

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Vasconcelos, LuisLehto, TõnisHällbrink, MattiasLangel, Ülo
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