Synthesis and evaluation of fluorescent Pam3Cys peptide conjugates

Geoffroy P.P. Gential, Nataschja I. Ho, Fabrizio Chiodo, Nico Meeuwenoord, Ferry Ossendorp, Herman S. Overkleeft, Gijs A. van der Marel, Dmitri V. Filippov

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

Chirally pure R- and S-epimers of TLR2 ligand Pam3CysSK4were prepared and separately conjugated to an OVA model epitope, in which lysine was replaced by azidonorleucine. The azide function in the conjugate permitted labelling with different fluorophores by use of strain-promoted 3+2 cycloaddition. The R-epimer of the labelled conjugates induced TLR2-dependent DC maturation, while S-epimer proved to be inactive. Combining the lipophilicity of Pam3CysSK4ligand with fluorophores influenced the solubility of the resulting conjugates in an unpredictable way and only the conjugates labelled with Cy-5 were suitable for confocal fluorescence microscopy experiments. It was shown that both epimers of the Cy-5 labelled lipopeptides were internalized equally well, indicating TLR2-independent cellular uptake. The presented results demonstrate the usefulness of strain-promoted azide-alkyne cycloaddition in the labelling of highly lipophilic lipopeptides without disturbing the in vitro activity of these conjugates with respect to activation of TLR-2.

Original languageEnglish
Pages (from-to)3641-3645
Number of pages5
JournalBioorganic and Medicinal Chemistry Letters
Volume26
Issue number15
DOIs
Publication statusPublished - 1 Jan 2016

Cite this

Gential, G. P. P., Ho, N. I., Chiodo, F., Meeuwenoord, N., Ossendorp, F., Overkleeft, H. S., ... Filippov, D. V. (2016). Synthesis and evaluation of fluorescent Pam3Cys peptide conjugates. Bioorganic and Medicinal Chemistry Letters, 26(15), 3641-3645. https://doi.org/10.1016/j.bmcl.2016.05.094
Gential, Geoffroy P.P. ; Ho, Nataschja I. ; Chiodo, Fabrizio ; Meeuwenoord, Nico ; Ossendorp, Ferry ; Overkleeft, Herman S. ; van der Marel, Gijs A. ; Filippov, Dmitri V. / Synthesis and evaluation of fluorescent Pam3Cys peptide conjugates. In: Bioorganic and Medicinal Chemistry Letters. 2016 ; Vol. 26, No. 15. pp. 3641-3645.
@article{d247afbad9ca49a08ace9dc49496f968,
title = "Synthesis and evaluation of fluorescent Pam3Cys peptide conjugates",
abstract = "Chirally pure R- and S-epimers of TLR2 ligand Pam3CysSK4were prepared and separately conjugated to an OVA model epitope, in which lysine was replaced by azidonorleucine. The azide function in the conjugate permitted labelling with different fluorophores by use of strain-promoted 3+2 cycloaddition. The R-epimer of the labelled conjugates induced TLR2-dependent DC maturation, while S-epimer proved to be inactive. Combining the lipophilicity of Pam3CysSK4ligand with fluorophores influenced the solubility of the resulting conjugates in an unpredictable way and only the conjugates labelled with Cy-5 were suitable for confocal fluorescence microscopy experiments. It was shown that both epimers of the Cy-5 labelled lipopeptides were internalized equally well, indicating TLR2-independent cellular uptake. The presented results demonstrate the usefulness of strain-promoted azide-alkyne cycloaddition in the labelling of highly lipophilic lipopeptides without disturbing the in vitro activity of these conjugates with respect to activation of TLR-2.",
keywords = "Fluorescent labeling, Lipopeptides, Strain promoted cycloaddition, Toll-like receptor 2",
author = "Gential, {Geoffroy P.P.} and Ho, {Nataschja I.} and Fabrizio Chiodo and Nico Meeuwenoord and Ferry Ossendorp and Overkleeft, {Herman S.} and {van der Marel}, {Gijs A.} and Filippov, {Dmitri V.}",
year = "2016",
month = "1",
day = "1",
doi = "10.1016/j.bmcl.2016.05.094",
language = "English",
volume = "26",
pages = "3641--3645",
journal = "Bioorganic and Medicinal Chemistry Letters",
issn = "0960-894X",
publisher = "Elsevier Limited",
number = "15",

}

Gential, GPP, Ho, NI, Chiodo, F, Meeuwenoord, N, Ossendorp, F, Overkleeft, HS, van der Marel, GA & Filippov, DV 2016, 'Synthesis and evaluation of fluorescent Pam3Cys peptide conjugates' Bioorganic and Medicinal Chemistry Letters, vol. 26, no. 15, pp. 3641-3645. https://doi.org/10.1016/j.bmcl.2016.05.094

Synthesis and evaluation of fluorescent Pam3Cys peptide conjugates. / Gential, Geoffroy P.P.; Ho, Nataschja I.; Chiodo, Fabrizio; Meeuwenoord, Nico; Ossendorp, Ferry; Overkleeft, Herman S.; van der Marel, Gijs A.; Filippov, Dmitri V.

In: Bioorganic and Medicinal Chemistry Letters, Vol. 26, No. 15, 01.01.2016, p. 3641-3645.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Synthesis and evaluation of fluorescent Pam3Cys peptide conjugates

AU - Gential, Geoffroy P.P.

AU - Ho, Nataschja I.

AU - Chiodo, Fabrizio

AU - Meeuwenoord, Nico

AU - Ossendorp, Ferry

AU - Overkleeft, Herman S.

AU - van der Marel, Gijs A.

AU - Filippov, Dmitri V.

PY - 2016/1/1

Y1 - 2016/1/1

N2 - Chirally pure R- and S-epimers of TLR2 ligand Pam3CysSK4were prepared and separately conjugated to an OVA model epitope, in which lysine was replaced by azidonorleucine. The azide function in the conjugate permitted labelling with different fluorophores by use of strain-promoted 3+2 cycloaddition. The R-epimer of the labelled conjugates induced TLR2-dependent DC maturation, while S-epimer proved to be inactive. Combining the lipophilicity of Pam3CysSK4ligand with fluorophores influenced the solubility of the resulting conjugates in an unpredictable way and only the conjugates labelled with Cy-5 were suitable for confocal fluorescence microscopy experiments. It was shown that both epimers of the Cy-5 labelled lipopeptides were internalized equally well, indicating TLR2-independent cellular uptake. The presented results demonstrate the usefulness of strain-promoted azide-alkyne cycloaddition in the labelling of highly lipophilic lipopeptides without disturbing the in vitro activity of these conjugates with respect to activation of TLR-2.

AB - Chirally pure R- and S-epimers of TLR2 ligand Pam3CysSK4were prepared and separately conjugated to an OVA model epitope, in which lysine was replaced by azidonorleucine. The azide function in the conjugate permitted labelling with different fluorophores by use of strain-promoted 3+2 cycloaddition. The R-epimer of the labelled conjugates induced TLR2-dependent DC maturation, while S-epimer proved to be inactive. Combining the lipophilicity of Pam3CysSK4ligand with fluorophores influenced the solubility of the resulting conjugates in an unpredictable way and only the conjugates labelled with Cy-5 were suitable for confocal fluorescence microscopy experiments. It was shown that both epimers of the Cy-5 labelled lipopeptides were internalized equally well, indicating TLR2-independent cellular uptake. The presented results demonstrate the usefulness of strain-promoted azide-alkyne cycloaddition in the labelling of highly lipophilic lipopeptides without disturbing the in vitro activity of these conjugates with respect to activation of TLR-2.

KW - Fluorescent labeling

KW - Lipopeptides

KW - Strain promoted cycloaddition

KW - Toll-like receptor 2

UR - http://www.scopus.com/inward/record.url?scp=84973484161&partnerID=8YFLogxK

U2 - 10.1016/j.bmcl.2016.05.094

DO - 10.1016/j.bmcl.2016.05.094

M3 - Article

VL - 26

SP - 3641

EP - 3645

JO - Bioorganic and Medicinal Chemistry Letters

JF - Bioorganic and Medicinal Chemistry Letters

SN - 0960-894X

IS - 15

ER -