Cargando…

In Vivo Behavior of the Antibacterial Peptide Cyclo[RRRWFW], Explored Using a 3-Hydroxychromone-Derived Fluorescent Amino Acid

Labeling biomolecules with fluorescent labels is an established tool for structural, biochemical, and biophysical studies; however, it remains underused for small peptides. In this work, an amino acid bearing a 3-hydroxychromone fluorophore, 2-amino-3-(2-(furan-2-yl)-3-hydroxy-4-oxo-4H-chromen-6-yl)...

Descripción completa

Detalles Bibliográficos
Autores principales: Afonin, Sergii, Koniev, Serhii, Préau, Laetitia, Takamiya, Masanari, Strizhak, Alexander V., Babii, Oleg, Hrebonkin, Andrii, Pivovarenko, Vasyl G., Dathe, Margitta, le Noble, Ferdinand, Rastegar, Sepand, Strähle, Uwe, Ulrich, Anne S., Komarov, Igor V.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8273159/
https://www.ncbi.nlm.nih.gov/pubmed/34262894
http://dx.doi.org/10.3389/fchem.2021.688446
_version_ 1783721311116197888
author Afonin, Sergii
Koniev, Serhii
Préau, Laetitia
Takamiya, Masanari
Strizhak, Alexander V.
Babii, Oleg
Hrebonkin, Andrii
Pivovarenko, Vasyl G.
Dathe, Margitta
le Noble, Ferdinand
Rastegar, Sepand
Strähle, Uwe
Ulrich, Anne S.
Komarov, Igor V.
author_facet Afonin, Sergii
Koniev, Serhii
Préau, Laetitia
Takamiya, Masanari
Strizhak, Alexander V.
Babii, Oleg
Hrebonkin, Andrii
Pivovarenko, Vasyl G.
Dathe, Margitta
le Noble, Ferdinand
Rastegar, Sepand
Strähle, Uwe
Ulrich, Anne S.
Komarov, Igor V.
author_sort Afonin, Sergii
collection PubMed
description Labeling biomolecules with fluorescent labels is an established tool for structural, biochemical, and biophysical studies; however, it remains underused for small peptides. In this work, an amino acid bearing a 3-hydroxychromone fluorophore, 2-amino-3-(2-(furan-2-yl)-3-hydroxy-4-oxo-4H-chromen-6-yl)propanoic acid (FHC), was incorporated in a known hexameric antimicrobial peptide, cyclo[RRRWFW] (cWFW), in place of aromatic residues. Circular dichroism spectropolarimetry and antibacterial activity measurements demonstrated that the FHC residue perturbs the peptide structure depending on labeling position but does not modify the activity of cWFW significantly. FHC thus can be considered an adequate label for studies of the parent peptide. Several analytical and imaging techniques were used to establish the activity of the obtained labeled cWFW analogues toward animal cells and to study the behavior of the peptides in a multicellular organism. The 3-hydroxychromone fluorophore can undergo excited-state intramolecular proton transfer (ESIPT), resulting in double-band emission from its two tautomeric forms. This feature allowed us to get insights into conformational equilibria of the labeled peptides, localize the cWFW analogues in human cells (HeLa and HEK293) and zebrafish embryos, and assess the polarity of the local environment around the label by confocal fluorescence microscopy. We found that the labeled peptides efficiently penetrated cancerous cells and localized mainly in lipid-containing and/or other nonpolar subcellular compartments. In the zebrafish embryo, the peptides remained in the bloodstream upon injection into the cardinal vein, presumably adhering to lipoproteins and/or microvesicles. They did not diffuse into any tissue to a significant extent during the first 3 h after administration. This study demonstrated the utility of fluorescent labeling by double-emission labels to evaluate biologically active peptides as potential drug candidates in vivo.
format Online
Article
Text
id pubmed-8273159
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher Frontiers Media S.A.
record_format MEDLINE/PubMed
spelling pubmed-82731592021-07-13 In Vivo Behavior of the Antibacterial Peptide Cyclo[RRRWFW], Explored Using a 3-Hydroxychromone-Derived Fluorescent Amino Acid Afonin, Sergii Koniev, Serhii Préau, Laetitia Takamiya, Masanari Strizhak, Alexander V. Babii, Oleg Hrebonkin, Andrii Pivovarenko, Vasyl G. Dathe, Margitta le Noble, Ferdinand Rastegar, Sepand Strähle, Uwe Ulrich, Anne S. Komarov, Igor V. Front Chem Chemistry Labeling biomolecules with fluorescent labels is an established tool for structural, biochemical, and biophysical studies; however, it remains underused for small peptides. In this work, an amino acid bearing a 3-hydroxychromone fluorophore, 2-amino-3-(2-(furan-2-yl)-3-hydroxy-4-oxo-4H-chromen-6-yl)propanoic acid (FHC), was incorporated in a known hexameric antimicrobial peptide, cyclo[RRRWFW] (cWFW), in place of aromatic residues. Circular dichroism spectropolarimetry and antibacterial activity measurements demonstrated that the FHC residue perturbs the peptide structure depending on labeling position but does not modify the activity of cWFW significantly. FHC thus can be considered an adequate label for studies of the parent peptide. Several analytical and imaging techniques were used to establish the activity of the obtained labeled cWFW analogues toward animal cells and to study the behavior of the peptides in a multicellular organism. The 3-hydroxychromone fluorophore can undergo excited-state intramolecular proton transfer (ESIPT), resulting in double-band emission from its two tautomeric forms. This feature allowed us to get insights into conformational equilibria of the labeled peptides, localize the cWFW analogues in human cells (HeLa and HEK293) and zebrafish embryos, and assess the polarity of the local environment around the label by confocal fluorescence microscopy. We found that the labeled peptides efficiently penetrated cancerous cells and localized mainly in lipid-containing and/or other nonpolar subcellular compartments. In the zebrafish embryo, the peptides remained in the bloodstream upon injection into the cardinal vein, presumably adhering to lipoproteins and/or microvesicles. They did not diffuse into any tissue to a significant extent during the first 3 h after administration. This study demonstrated the utility of fluorescent labeling by double-emission labels to evaluate biologically active peptides as potential drug candidates in vivo. Frontiers Media S.A. 2021-06-28 /pmc/articles/PMC8273159/ /pubmed/34262894 http://dx.doi.org/10.3389/fchem.2021.688446 Text en Copyright © 2021 Afonin, Koniev, Préau, Takamiya, Strizhak, Babii, Hrebonkin, Pivovarenko, Dathe, le Noble, Rastegar, Strähle, Ulrich and Komarov. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Chemistry
Afonin, Sergii
Koniev, Serhii
Préau, Laetitia
Takamiya, Masanari
Strizhak, Alexander V.
Babii, Oleg
Hrebonkin, Andrii
Pivovarenko, Vasyl G.
Dathe, Margitta
le Noble, Ferdinand
Rastegar, Sepand
Strähle, Uwe
Ulrich, Anne S.
Komarov, Igor V.
In Vivo Behavior of the Antibacterial Peptide Cyclo[RRRWFW], Explored Using a 3-Hydroxychromone-Derived Fluorescent Amino Acid
title In Vivo Behavior of the Antibacterial Peptide Cyclo[RRRWFW], Explored Using a 3-Hydroxychromone-Derived Fluorescent Amino Acid
title_full In Vivo Behavior of the Antibacterial Peptide Cyclo[RRRWFW], Explored Using a 3-Hydroxychromone-Derived Fluorescent Amino Acid
title_fullStr In Vivo Behavior of the Antibacterial Peptide Cyclo[RRRWFW], Explored Using a 3-Hydroxychromone-Derived Fluorescent Amino Acid
title_full_unstemmed In Vivo Behavior of the Antibacterial Peptide Cyclo[RRRWFW], Explored Using a 3-Hydroxychromone-Derived Fluorescent Amino Acid
title_short In Vivo Behavior of the Antibacterial Peptide Cyclo[RRRWFW], Explored Using a 3-Hydroxychromone-Derived Fluorescent Amino Acid
title_sort in vivo behavior of the antibacterial peptide cyclo[rrrwfw], explored using a 3-hydroxychromone-derived fluorescent amino acid
topic Chemistry
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8273159/
https://www.ncbi.nlm.nih.gov/pubmed/34262894
http://dx.doi.org/10.3389/fchem.2021.688446
work_keys_str_mv AT afoninsergii invivobehavioroftheantibacterialpeptidecyclorrrwfwexploredusinga3hydroxychromonederivedfluorescentaminoacid
AT konievserhii invivobehavioroftheantibacterialpeptidecyclorrrwfwexploredusinga3hydroxychromonederivedfluorescentaminoacid
AT preaulaetitia invivobehavioroftheantibacterialpeptidecyclorrrwfwexploredusinga3hydroxychromonederivedfluorescentaminoacid
AT takamiyamasanari invivobehavioroftheantibacterialpeptidecyclorrrwfwexploredusinga3hydroxychromonederivedfluorescentaminoacid
AT strizhakalexanderv invivobehavioroftheantibacterialpeptidecyclorrrwfwexploredusinga3hydroxychromonederivedfluorescentaminoacid
AT babiioleg invivobehavioroftheantibacterialpeptidecyclorrrwfwexploredusinga3hydroxychromonederivedfluorescentaminoacid
AT hrebonkinandrii invivobehavioroftheantibacterialpeptidecyclorrrwfwexploredusinga3hydroxychromonederivedfluorescentaminoacid
AT pivovarenkovasylg invivobehavioroftheantibacterialpeptidecyclorrrwfwexploredusinga3hydroxychromonederivedfluorescentaminoacid
AT dathemargitta invivobehavioroftheantibacterialpeptidecyclorrrwfwexploredusinga3hydroxychromonederivedfluorescentaminoacid
AT lenobleferdinand invivobehavioroftheantibacterialpeptidecyclorrrwfwexploredusinga3hydroxychromonederivedfluorescentaminoacid
AT rastegarsepand invivobehavioroftheantibacterialpeptidecyclorrrwfwexploredusinga3hydroxychromonederivedfluorescentaminoacid
AT strahleuwe invivobehavioroftheantibacterialpeptidecyclorrrwfwexploredusinga3hydroxychromonederivedfluorescentaminoacid
AT ulrichannes invivobehavioroftheantibacterialpeptidecyclorrrwfwexploredusinga3hydroxychromonederivedfluorescentaminoacid
AT komarovigorv invivobehavioroftheantibacterialpeptidecyclorrrwfwexploredusinga3hydroxychromonederivedfluorescentaminoacid