Crystal Structure of Escherichia coli originated MCR-1, a phosphoethanolamine transferase for Colistin Resistance

Hu, Maogui; Guo, Jiubiao; Cheng, Qipeng; Yang, Zhiqiang; Chan, Edward Wai Chi; Chen, Sheng; Hao, Quan

doi:10.1038/srep38793

Cited by 69 publications

(66 citation statements)

References 14 publications

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“…Notably, Thr315 in the ICR Mc cat active site occupies the position corresponding to the phosphate-accepting nucleophilic residue in the other PEtN transferases. 26 , 27 , 29–32 Analysis of ICR Mc cat residues interacting with both Zn 2+ and the phosphate ions also highlights His429 as a functionally important residue, although its interaction with the Zn 2+ was mediated through a hydrogen bond with the coordinated water molecule. In addition to the Zn 2+ and phosphate ions, the ICR Mc cat active site contained a sulfate ion positioned at 11 Å distance from the phosphate ion described above.…”

Section: Resultsmentioning

confidence: 99%

Substrate Recognition by a Colistin Resistance Enzyme from Moraxella catarrhalis

et al. 2018

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Lipid A phosphoethanolamine (PEtN) transferases render bacteria resistant to the last resort antibiotic colistin. The recent discoveries of pathogenic bacteria harboring plasmid-borne PEtN transferase (mcr) genes have illustrated the serious potential for wide dissemination of these resistance elements. The origin of mcr-1 is traced to Moraxella species co-occupying environmental niches with Enterobacteriaceae. Here, we describe the crystal structure of the catalytic domain of the chromosomally encoded colistin resistance PEtN transferase, ICRMc (for intrinsic colistin resistance) of Moraxella catarrhalis. The ICRMc structure in complex with PEtN reveals key molecular details including specific residues involved in catalysis and PEtN binding. It also demonstrates that ICRMc catalytic domain dimerization is required for substrate binding. Our structure-guided phylogenetic analysis provides sequence signatures defining potentially colistin-active representatives in this enzyme family. Combined, these results advance the molecular and mechanistic understanding of PEtN transferases and illuminate their origins.

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Section: Resultsmentioning

confidence: 99%

Substrate Recognition by a Colistin Resistance Enzyme from Moraxella catarrhalis

et al. 2018

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“…2). The functionally characterized transferases included three EptA homologs for which the catalytic domain structure has been resolved, namely, EptA Nm (LptA) (15) and the plasmid-mediated Mcr-1, both of which confer resistance to polymyxins in certain Gram-negative bacteria via the addi-tion of PEtn to lipid A (16,17), and Campylobacter jejuni protein EptC (18), a multifunctional transferase that can add PEtn to a range of cell components, including lipid A, heptose within the LPS inner core, and flagella (19)(20)(21). The amino acid sequences of these proteins, their active sites, and the associated metal binding sites are highly conserved.…”

Section: Resultsmentioning

confidence: 99%

Characterization of Two Novel Lipopolysaccharide Phosphoethanolamine Transferases in Pasteurella multocida and Their Role in Resistance to Cathelicidin-2

et al. 2017

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The lipopolysaccharide (LPS) produced by the Gram-negative bacterial pathogen Pasteurella multocida has phosphoethanolamine (PEtn) residues attached to lipid A, 3-deoxy-D-manno-octulosonic acid (Kdo), heptose, and galactose. In this report, we show that PEtn is transferred to lipid A by the P. multocida EptA homologue, PetL, and is transferred to galactose by a novel PEtn transferase that is unique to P. multocida called PetG. Transcriptomic analyses indicated that petL expression was positively regulated by the global regulator Fis and negatively regulated by an Hfq-dependent small RNA. Importantly, we have identified a novel PEtn transferase called PetK that is responsible for PEtn addition to the single Kdo molecule (Kdo 1 ), directly linked to lipid A in the P. multocida glycoform A LPS. In vitro assays showed that the presence of a functional petL and petK, and therefore the presence of PEtn on lipid A and Kdo 1 , was essential for resistance to the cationic, antimicrobial peptide cathelicidin-2. The importance of PEtn on Kdo 1 and the identification of the transferase responsible for this addition have not previously been shown. Phylogenetic analysis revealed that PetK is the first representative of a new family of predicted PEtn transferases. The PetK family consists of uncharacterized proteins from a range of Gram-negative bacteria that produce LPS glycoforms with only one Kdo molecule, including pathogenic species within the genera Vibrio, Bordetella, and Haemophilus. We predict that many of these bacteria will require the addition of PEtn to Kdo for maximum protection against host antimicrobial peptides.

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“…To date, most of the research in this field has focused on epidemiologic issues, and several independent research groups have obtained the crystal structure of the MCR-1 catalytic domain (cMCR-1) alone (30)(31)(32)(33). Although the detailed molecular mechanism of its substrate binding remains poorly understood, it may provide an important way to combat antibiotic resistance, such as with specific MCR-1 inhibitors.…”

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confidence: 99%

Substrate analog interaction with MCR‐1 offers insight into the rising threat of the plasmid‐mediated transferable colistin resistance

et al. 2018

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Colistin is considered a last-resort antibiotic against most gram-negative bacteria. Recent discoveries of a plasmid-mediated, transferable mobilized colistin-resistance gene ( mcr-1) on all continents have heralded the imminent emergence of pan-drug-resistant superbacteria. The inner-membrane protein MCR-1 can catalyze the transfer of phosphoethanolamine (PEA) to lipid A, resulting in colistin resistance. However, little is known about the mechanism, and few drugs exist to address this issue. We present crystal structures revealing the MCR-1 catalytic domain (cMCR-1) as a monozinc metalloprotein with ethanolamine (ETA) and d-glucose, respectively, thus highlighting 2 possible substrate-binding pockets in the MCR-1-catalyzed PEA transfer reaction. Mutation of the residues involved in ETA and d-glucose binding impairs colistin resistance in recombinant Escherichia coli containing full-length MCR-1. Partial analogs of the substrate are used for cocrystallization with cMCR-1, providing valuable information about the family of PEA transferases. One of the analogs, ETA, causes clear inhibition of polymyxin B resistance, highlighting its potential for drug development. These data demonstrate the crucial role of the PEA- and lipid A-binding pockets and provide novel insights into the structure-based mechanisms, important drug-target hot spots, and a drug template for further drug development to combat the urgent, rising threat of MCR-1-mediated antibiotic resistance.-Wei, P., Song, G., Shi, M., Zhou, Y., Liu, Y., Lei, J., Chen, P., Yin, L. Substrate analog interaction with MCR-1 offers insight into the rising threat of the plasmid-mediated transferable colistin resistance.

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Crystal Structure of Escherichia coli originated MCR-1, a phosphoethanolamine transferase for Colistin Resistance

Cited by 69 publications

References 14 publications

Substrate Recognition by a Colistin Resistance Enzyme from Moraxella catarrhalis

Substrate Recognition by a Colistin Resistance Enzyme from Moraxella catarrhalis

Characterization of Two Novel Lipopolysaccharide Phosphoethanolamine Transferases in Pasteurella multocida and Their Role in Resistance to Cathelicidin-2

Substrate analog interaction with MCR‐1 offers insight into the rising threat of the plasmid‐mediated transferable colistin resistance

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