@article{Chmelova2023-09-15Multi-67703,
  year={2023},
  doi={10.1002/pro.4751},
  title={Multimeric structure of a subfamily III haloalkane dehalogenase-like enzyme solved by combination of cryo-EM and X-ray crystallography},
  number={10},
  volume={32},
  issn={0961-8368},
  journal={Protein Science},
  author={Chmelova, Klaudia and Gao, Tadeja and Polak, Martin and Schenkmayerova, Andrea and Croll, Tristan I. and Shaikh, Tanvir R. and Skarupova, Jana and Chaloupkova, Radka and Diederichs, Kay and Marek, Martin},
  note={Article Number: e4751}
}

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    <dc:contributor>Croll, Tristan I.</dc:contributor>
    <dc:contributor>Schenkmayerova, Andrea</dc:contributor>
    <dc:creator>Chaloupkova, Radka</dc:creator>
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    <dc:contributor>Chmelova, Klaudia</dc:contributor>
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    <dc:contributor>Diederichs, Kay</dc:contributor>
    <dc:creator>Diederichs, Kay</dc:creator>
    <dc:creator>Shaikh, Tanvir R.</dc:creator>
    <dcterms:title>Multimeric structure of a subfamily III haloalkane dehalogenase-like enzyme solved by combination of cryo-EM and X-ray crystallography</dcterms:title>
    <dc:contributor>Gao, Tadeja</dc:contributor>
    <dcterms:available rdf:datatype="http://www.w3.org/2001/XMLSchema#dateTime">2023-08-30T08:51:27Z</dcterms:available>
    <dc:creator>Skarupova, Jana</dc:creator>
    <dc:creator>Chmelova, Klaudia</dc:creator>
    <dspace:isPartOfCollection rdf:resource="https://kops.uni-konstanz.de/server/rdf/resource/123456789/28"/>
    <dc:creator>Polak, Martin</dc:creator>
    <dc:creator>Croll, Tristan I.</dc:creator>
    <dc:contributor>Chaloupkova, Radka</dc:contributor>
    <dc:contributor>Skarupova, Jana</dc:contributor>
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    <dcterms:issued>2023-09-15</dcterms:issued>
    <dc:creator>Schenkmayerova, Andrea</dc:creator>
    <dc:contributor>Shaikh, Tanvir R.</dc:contributor>
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    <dc:language>eng</dc:language>
    <dc:creator>Gao, Tadeja</dc:creator>
    <dcterms:abstract>Haloalkane dehalogenase (HLD) enzymes employ an S&lt;sub&gt;N&lt;/sub&gt;2 nucleophilic substitution mechanism to erase halogen substituents in diverse organohalogen compounds. Subfamily I and II HLDs are well-characterized enzymes, but a mode and purpose of multimerization of subfamily III HLDs are unknown. Here we probe the structural organization of DhmeA, a subfamily III HLD-like enzyme from the archaeon Haloferax mediterranei, by combining cryo-electron microscopy (cryo-EM) and X-ray crystallography. We show that full-length wild-type DhmeA forms diverse quaternary structures, ranging from small oligomers to large supramolecular ring-like assemblies of various sizes and symmetries. We optimized sample preparation steps, enabling three-dimensional reconstructions of an oligomeric species by single-particle cryo-EM. Moreover, we engineered a crystallizable mutant (DhmeA&lt;sup&gt;ΔGG&lt;/sup&gt;) that provided diffraction-quality crystals. The 3.3 Å crystal structure reveals that DhmeA&lt;sup&gt;ΔGG&lt;/sup&gt; forms a ring-like 20-mer structure with outer and inner diameter of ~200 Å and ~80 Å, respectively. An enzyme homodimer represents a basic repeating building unit of the crystallographic ring. Three assembly interfaces (dimerization, tetramerization and multimerization) were identified to form the supramolecular ring that displays a negatively charged exterior, while its interior part harboring catalytic sites is positively charged. Localization and exposure of catalytic machineries suggest a possible processing of large negatively charged macromolecular substrates.</dcterms:abstract>
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    <dc:contributor>Marek, Martin</dc:contributor>
    <dc:date rdf:datatype="http://www.w3.org/2001/XMLSchema#dateTime">2023-08-30T08:51:27Z</dc:date>
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    <dc:creator>Marek, Martin</dc:creator>
    <dc:contributor>Polak, Martin</dc:contributor>
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