Dodecapeptide Cathelicidins of Cetartiodactyla: Structure, Mechanism of Antimicrobial Action, and Synergistic Interaction With Other Cathelicidins

In this study, dodecapeptide cathelicidins were shown to be widespread antimicrobial peptides among the Cetruminantia clade. In particular, we investigated the dodecapeptide from the domestic goat Capra hircus, designated as ChDode and its unique ortholog from the sperm whale Physeter catodon (PcDod...

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Main Authors: Ilia A. Bolosov, Pavel V. Panteleev, Sergei V. Sychev, Stanislav V. Sukhanov, Pavel A. Mironov, Mikhail Yu. Myshkin, Zakhar O. Shenkarev, Tatiana V. Ovchinnikova
Format: Article
Language:English
Published: Frontiers Media S.A. 2021-08-01
Series:Frontiers in Microbiology
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Online Access:https://www.frontiersin.org/articles/10.3389/fmicb.2021.725526/full
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author Ilia A. Bolosov
Pavel V. Panteleev
Pavel V. Panteleev
Sergei V. Sychev
Stanislav V. Sukhanov
Pavel A. Mironov
Pavel A. Mironov
Mikhail Yu. Myshkin
Zakhar O. Shenkarev
Zakhar O. Shenkarev
Tatiana V. Ovchinnikova
Tatiana V. Ovchinnikova
Tatiana V. Ovchinnikova
author_facet Ilia A. Bolosov
Pavel V. Panteleev
Pavel V. Panteleev
Sergei V. Sychev
Stanislav V. Sukhanov
Pavel A. Mironov
Pavel A. Mironov
Mikhail Yu. Myshkin
Zakhar O. Shenkarev
Zakhar O. Shenkarev
Tatiana V. Ovchinnikova
Tatiana V. Ovchinnikova
Tatiana V. Ovchinnikova
author_sort Ilia A. Bolosov
collection DOAJ
description In this study, dodecapeptide cathelicidins were shown to be widespread antimicrobial peptides among the Cetruminantia clade. In particular, we investigated the dodecapeptide from the domestic goat Capra hircus, designated as ChDode and its unique ortholog from the sperm whale Physeter catodon (PcDode). ChDode contains two cysteine residues, while PcDode consists of two dodecapeptide building blocks and contains four cysteine residues. The recombinant analogs of the peptides were obtained by heterologous expression in Escherichia coli cells. The structures of the peptides were studied by circular dichroism (CD), FTIR, and NMR spectroscopy. It was demonstrated that PcDode adopts a β-hairpin structure in water and resembles β-hairpin antimicrobial peptides, while ChDode forms a β-structural antiparallel covalent dimer, stabilized by two intermonomer disulfide bonds. Both peptides reveal a significant right-handed twist about 200 degrees per 8 residues. In DPC micelles ChDode forms flat β-structural tetramers by antiparallel non-covalent association of the dimers. The tetramers incorporate into the micelles in transmembrane orientation. Incorporation into the micelles and dimerization significantly diminished the amplitude of backbone motions of ChDode at the picosecond-nanosecond timescale. When interacting with negatively charged membranes containing phosphatidylethanolamine (PE) and phosphatidylglycerol (PG), the ChDode peptide adopted similar oligomeric structure and was capable to form ion-conducting pores without membrane lysis. Despite modest antibacterial activity of ChDode, a considerable synergistic effect of this peptide in combination with another goat cathelicidin – the α-helical peptide ChMAP-28 was observed. This effect is based on an increase in permeability of bacterial membranes. In turn, this mechanism can lead to an increase in the efficiency of the combined action of the synergistic pair ChMAP-28 with the Pro-rich peptide mini-ChBac7.5Nα targeting the bacterial ribosome.
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spelling doaj.art-5bdff750a9d0479a9f0fb4bd9d62559d2022-12-21T19:58:09ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2021-08-011210.3389/fmicb.2021.725526725526Dodecapeptide Cathelicidins of Cetartiodactyla: Structure, Mechanism of Antimicrobial Action, and Synergistic Interaction With Other CathelicidinsIlia A. Bolosov0Pavel V. Panteleev1Pavel V. Panteleev2Sergei V. Sychev3Stanislav V. Sukhanov4Pavel A. Mironov5Pavel A. Mironov6Mikhail Yu. Myshkin7Zakhar O. Shenkarev8Zakhar O. Shenkarev9Tatiana V. Ovchinnikova10Tatiana V. Ovchinnikova11Tatiana V. Ovchinnikova12M. M. Shemyakin and Yu. A. Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, RussiaM. M. Shemyakin and Yu. A. Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, RussiaPhystech School of Biological and Medical Physics, Moscow Institute of Physics and Technology (State University), Dolgoprudny, RussiaM. M. Shemyakin and Yu. A. Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, RussiaM. M. Shemyakin and Yu. A. Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, RussiaM. M. Shemyakin and Yu. A. Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, RussiaFaculty of Biology, Lomonosov Moscow State University, Moscow, RussiaM. M. Shemyakin and Yu. A. Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, RussiaM. M. Shemyakin and Yu. A. Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, RussiaPhystech School of Biological and Medical Physics, Moscow Institute of Physics and Technology (State University), Dolgoprudny, RussiaM. M. Shemyakin and Yu. A. Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, RussiaPhystech School of Biological and Medical Physics, Moscow Institute of Physics and Technology (State University), Dolgoprudny, RussiaFaculty of Biology, Lomonosov Moscow State University, Moscow, RussiaIn this study, dodecapeptide cathelicidins were shown to be widespread antimicrobial peptides among the Cetruminantia clade. In particular, we investigated the dodecapeptide from the domestic goat Capra hircus, designated as ChDode and its unique ortholog from the sperm whale Physeter catodon (PcDode). ChDode contains two cysteine residues, while PcDode consists of two dodecapeptide building blocks and contains four cysteine residues. The recombinant analogs of the peptides were obtained by heterologous expression in Escherichia coli cells. The structures of the peptides were studied by circular dichroism (CD), FTIR, and NMR spectroscopy. It was demonstrated that PcDode adopts a β-hairpin structure in water and resembles β-hairpin antimicrobial peptides, while ChDode forms a β-structural antiparallel covalent dimer, stabilized by two intermonomer disulfide bonds. Both peptides reveal a significant right-handed twist about 200 degrees per 8 residues. In DPC micelles ChDode forms flat β-structural tetramers by antiparallel non-covalent association of the dimers. The tetramers incorporate into the micelles in transmembrane orientation. Incorporation into the micelles and dimerization significantly diminished the amplitude of backbone motions of ChDode at the picosecond-nanosecond timescale. When interacting with negatively charged membranes containing phosphatidylethanolamine (PE) and phosphatidylglycerol (PG), the ChDode peptide adopted similar oligomeric structure and was capable to form ion-conducting pores without membrane lysis. Despite modest antibacterial activity of ChDode, a considerable synergistic effect of this peptide in combination with another goat cathelicidin – the α-helical peptide ChMAP-28 was observed. This effect is based on an increase in permeability of bacterial membranes. In turn, this mechanism can lead to an increase in the efficiency of the combined action of the synergistic pair ChMAP-28 with the Pro-rich peptide mini-ChBac7.5Nα targeting the bacterial ribosome.https://www.frontiersin.org/articles/10.3389/fmicb.2021.725526/fullantimicrobial peptidecathelicidindodecapeptidegoatsynergyNMR
spellingShingle Ilia A. Bolosov
Pavel V. Panteleev
Pavel V. Panteleev
Sergei V. Sychev
Stanislav V. Sukhanov
Pavel A. Mironov
Pavel A. Mironov
Mikhail Yu. Myshkin
Zakhar O. Shenkarev
Zakhar O. Shenkarev
Tatiana V. Ovchinnikova
Tatiana V. Ovchinnikova
Tatiana V. Ovchinnikova
Dodecapeptide Cathelicidins of Cetartiodactyla: Structure, Mechanism of Antimicrobial Action, and Synergistic Interaction With Other Cathelicidins
Frontiers in Microbiology
antimicrobial peptide
cathelicidin
dodecapeptide
goat
synergy
NMR
title Dodecapeptide Cathelicidins of Cetartiodactyla: Structure, Mechanism of Antimicrobial Action, and Synergistic Interaction With Other Cathelicidins
title_full Dodecapeptide Cathelicidins of Cetartiodactyla: Structure, Mechanism of Antimicrobial Action, and Synergistic Interaction With Other Cathelicidins
title_fullStr Dodecapeptide Cathelicidins of Cetartiodactyla: Structure, Mechanism of Antimicrobial Action, and Synergistic Interaction With Other Cathelicidins
title_full_unstemmed Dodecapeptide Cathelicidins of Cetartiodactyla: Structure, Mechanism of Antimicrobial Action, and Synergistic Interaction With Other Cathelicidins
title_short Dodecapeptide Cathelicidins of Cetartiodactyla: Structure, Mechanism of Antimicrobial Action, and Synergistic Interaction With Other Cathelicidins
title_sort dodecapeptide cathelicidins of cetartiodactyla structure mechanism of antimicrobial action and synergistic interaction with other cathelicidins
topic antimicrobial peptide
cathelicidin
dodecapeptide
goat
synergy
NMR
url https://www.frontiersin.org/articles/10.3389/fmicb.2021.725526/full
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