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Identification of Functional Interactome of Colistin Resistance Protein MCR-1 in Escherichia coli

The emergence and worldwide dissemination of plasmid-mediated colistin resistance gene mcr-1 has attracted global attention. The MCR-1 enzyme mediated colistin resistance by catalyzing phosphoethanolamine (PEA) transfer onto bacterial lipid A. However, the interaction partners of MCR-1 located in me...

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Autores principales: Li, Hui, Wang, Yingyu, Chen, Qiyan, Xia, Xi, Shen, Jianzhong, Wang, Yang, Shao, Bing
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7868338/
https://www.ncbi.nlm.nih.gov/pubmed/33569043
http://dx.doi.org/10.3389/fmicb.2020.583185
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author Li, Hui
Wang, Yingyu
Chen, Qiyan
Xia, Xi
Shen, Jianzhong
Wang, Yang
Shao, Bing
author_facet Li, Hui
Wang, Yingyu
Chen, Qiyan
Xia, Xi
Shen, Jianzhong
Wang, Yang
Shao, Bing
author_sort Li, Hui
collection PubMed
description The emergence and worldwide dissemination of plasmid-mediated colistin resistance gene mcr-1 has attracted global attention. The MCR-1 enzyme mediated colistin resistance by catalyzing phosphoethanolamine (PEA) transfer onto bacterial lipid A. However, the interaction partners of MCR-1 located in membrane protein in E. coli are unknown. Co-immunoprecipitation (Co-IP) and Mass Spectrometry were performed to define the interacting proteins of MCR-1. A total of three different anti-MCR-1 monoclonal antibody (mAbs) were prepared and 3G4 mAb was selected as the bait protein by compared their suitability for Co-IP. We identified 53, 13, and 14 interacting proteins in E. coli BL21 (DE3) (pET28a-mcr-1), E. coli BL21 (DE3) (pET28a-mcr-1-200), and E. coli DH5α (pUC19-mcr-1), respectively. Six proteins, including the stress response proteins DnaK (chaperone protein) and SspB (stringent starvation protein B), the transcriptional regulation protein H-NS, and ribosomal proteins (RpsE, RpsJ, and RpsP) were identified in all these three strains. These MCR-1-interacting proteins were mainly involved in ribosome and RNA degradation, suggesting that MCR-1 influences the protein biosynthesis through the interaction with ribosomal protein. Multidrug efflux pump AcrA and TolC were important interacting membrane proteins of MCR-1 referred to drug efflux during the PEA modification of the bacterial cell membrane. Overall, we firstly identified the functional interactome profile of MCR-1 in E. coli and discovered that two-component AcrA-TolC multidrug efflux pump was involved in mcr-1-mediated colistin resistance.
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spelling pubmed-78683382021-02-09 Identification of Functional Interactome of Colistin Resistance Protein MCR-1 in Escherichia coli Li, Hui Wang, Yingyu Chen, Qiyan Xia, Xi Shen, Jianzhong Wang, Yang Shao, Bing Front Microbiol Microbiology The emergence and worldwide dissemination of plasmid-mediated colistin resistance gene mcr-1 has attracted global attention. The MCR-1 enzyme mediated colistin resistance by catalyzing phosphoethanolamine (PEA) transfer onto bacterial lipid A. However, the interaction partners of MCR-1 located in membrane protein in E. coli are unknown. Co-immunoprecipitation (Co-IP) and Mass Spectrometry were performed to define the interacting proteins of MCR-1. A total of three different anti-MCR-1 monoclonal antibody (mAbs) were prepared and 3G4 mAb was selected as the bait protein by compared their suitability for Co-IP. We identified 53, 13, and 14 interacting proteins in E. coli BL21 (DE3) (pET28a-mcr-1), E. coli BL21 (DE3) (pET28a-mcr-1-200), and E. coli DH5α (pUC19-mcr-1), respectively. Six proteins, including the stress response proteins DnaK (chaperone protein) and SspB (stringent starvation protein B), the transcriptional regulation protein H-NS, and ribosomal proteins (RpsE, RpsJ, and RpsP) were identified in all these three strains. These MCR-1-interacting proteins were mainly involved in ribosome and RNA degradation, suggesting that MCR-1 influences the protein biosynthesis through the interaction with ribosomal protein. Multidrug efflux pump AcrA and TolC were important interacting membrane proteins of MCR-1 referred to drug efflux during the PEA modification of the bacterial cell membrane. Overall, we firstly identified the functional interactome profile of MCR-1 in E. coli and discovered that two-component AcrA-TolC multidrug efflux pump was involved in mcr-1-mediated colistin resistance. Frontiers Media S.A. 2021-01-25 /pmc/articles/PMC7868338/ /pubmed/33569043 http://dx.doi.org/10.3389/fmicb.2020.583185 Text en Copyright © 2021 Li, Wang, Chen, Xia, Shen, Wang and Shao. http://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 Microbiology
Li, Hui
Wang, Yingyu
Chen, Qiyan
Xia, Xi
Shen, Jianzhong
Wang, Yang
Shao, Bing
Identification of Functional Interactome of Colistin Resistance Protein MCR-1 in Escherichia coli
title Identification of Functional Interactome of Colistin Resistance Protein MCR-1 in Escherichia coli
title_full Identification of Functional Interactome of Colistin Resistance Protein MCR-1 in Escherichia coli
title_fullStr Identification of Functional Interactome of Colistin Resistance Protein MCR-1 in Escherichia coli
title_full_unstemmed Identification of Functional Interactome of Colistin Resistance Protein MCR-1 in Escherichia coli
title_short Identification of Functional Interactome of Colistin Resistance Protein MCR-1 in Escherichia coli
title_sort identification of functional interactome of colistin resistance protein mcr-1 in escherichia coli
topic Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7868338/
https://www.ncbi.nlm.nih.gov/pubmed/33569043
http://dx.doi.org/10.3389/fmicb.2020.583185
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