A Biochemical Deconstruction-Based Strategy to Assist the Characterization of Bacterial Electric Conductive Filaments
Periplasmic nanowires and electric conductive filaments made of the polymeric assembly of <i>c</i>-type cytochromes from <i>Geobacter sulfurreducens</i> bacterium are crucial for electron storage and/or extracellular electron transfer. The elucidation of the redox properties...
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-04-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/24/8/7032 |
| _version_ | 1797605171682869248 |
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| author | Marta A. Silva Ana P. Fernandes David L. Turner Carlos A. Salgueiro |
| author_facet | Marta A. Silva Ana P. Fernandes David L. Turner Carlos A. Salgueiro |
| author_sort | Marta A. Silva |
| collection | DOAJ |
| description | Periplasmic nanowires and electric conductive filaments made of the polymeric assembly of <i>c</i>-type cytochromes from <i>Geobacter sulfurreducens</i> bacterium are crucial for electron storage and/or extracellular electron transfer. The elucidation of the redox properties of each heme is fundamental to the understanding of the electron transfer mechanisms in these systems, which first requires the specific assignment of the heme NMR signals. The high number of hemes and the molecular weight of the nanowires dramatically decrease the spectral resolution and make this assignment extremely complex or unattainable. The nanowire cytochrome GSU1996 (~42 kDa) is composed of four domains (A to D) each containing three <i>c</i>-type heme groups. In this work, the individual domains (A to D), bi-domains (AB, CD) and full-length nanowire were separately produced at natural abundance. Sufficient protein expression was obtained for domains C (~11 kDa/three hemes) and D (~10 kDa/three hemes), as well as for bi-domain CD (~21 kDa/six hemes). Using 2D-NMR experiments, the assignment of the heme proton NMR signals for domains C and D was obtained and then used to guide the assignment of the corresponding signals in the hexaheme bi-domain CD. This new biochemical deconstruction-based procedure, using nanowire GSU1996 as a model, establishes a new strategy to functionally characterize large multiheme cytochromes. |
| first_indexed | 2024-03-11T04:57:20Z |
| format | Article |
| id | doaj.art-a38e959d6a704019bdeaf0514dbab672 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-11T04:57:20Z |
| publishDate | 2023-04-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-a38e959d6a704019bdeaf0514dbab6722023-11-17T19:34:11ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-04-01248703210.3390/ijms24087032A Biochemical Deconstruction-Based Strategy to Assist the Characterization of Bacterial Electric Conductive FilamentsMarta A. Silva0Ana P. Fernandes1David L. Turner2Carlos A. Salgueiro3Associate Laboratory, i4HB—Institute for Health and Bioeconomy, School of Science and Technology, NOVA University Lisbon, 2819-516 Caparica, PortugalAssociate Laboratory, i4HB—Institute for Health and Bioeconomy, School of Science and Technology, NOVA University Lisbon, 2819-516 Caparica, PortugalInstituto de Tecnologia Química e Biológica António Xavier, NOVA University Lisbon, 2780-157 Oeiras, PortugalAssociate Laboratory, i4HB—Institute for Health and Bioeconomy, School of Science and Technology, NOVA University Lisbon, 2819-516 Caparica, PortugalPeriplasmic nanowires and electric conductive filaments made of the polymeric assembly of <i>c</i>-type cytochromes from <i>Geobacter sulfurreducens</i> bacterium are crucial for electron storage and/or extracellular electron transfer. The elucidation of the redox properties of each heme is fundamental to the understanding of the electron transfer mechanisms in these systems, which first requires the specific assignment of the heme NMR signals. The high number of hemes and the molecular weight of the nanowires dramatically decrease the spectral resolution and make this assignment extremely complex or unattainable. The nanowire cytochrome GSU1996 (~42 kDa) is composed of four domains (A to D) each containing three <i>c</i>-type heme groups. In this work, the individual domains (A to D), bi-domains (AB, CD) and full-length nanowire were separately produced at natural abundance. Sufficient protein expression was obtained for domains C (~11 kDa/three hemes) and D (~10 kDa/three hemes), as well as for bi-domain CD (~21 kDa/six hemes). Using 2D-NMR experiments, the assignment of the heme proton NMR signals for domains C and D was obtained and then used to guide the assignment of the corresponding signals in the hexaheme bi-domain CD. This new biochemical deconstruction-based procedure, using nanowire GSU1996 as a model, establishes a new strategy to functionally characterize large multiheme cytochromes.https://www.mdpi.com/1422-0067/24/8/7032<i>Geobacter</i>nanowiresmultiheme cytochromeselectron transferNMR |
| spellingShingle | Marta A. Silva Ana P. Fernandes David L. Turner Carlos A. Salgueiro A Biochemical Deconstruction-Based Strategy to Assist the Characterization of Bacterial Electric Conductive Filaments International Journal of Molecular Sciences <i>Geobacter</i> nanowires multiheme cytochromes electron transfer NMR |
| title | A Biochemical Deconstruction-Based Strategy to Assist the Characterization of Bacterial Electric Conductive Filaments |
| title_full | A Biochemical Deconstruction-Based Strategy to Assist the Characterization of Bacterial Electric Conductive Filaments |
| title_fullStr | A Biochemical Deconstruction-Based Strategy to Assist the Characterization of Bacterial Electric Conductive Filaments |
| title_full_unstemmed | A Biochemical Deconstruction-Based Strategy to Assist the Characterization of Bacterial Electric Conductive Filaments |
| title_short | A Biochemical Deconstruction-Based Strategy to Assist the Characterization of Bacterial Electric Conductive Filaments |
| title_sort | biochemical deconstruction based strategy to assist the characterization of bacterial electric conductive filaments |
| topic | <i>Geobacter</i> nanowires multiheme cytochromes electron transfer NMR |
| url | https://www.mdpi.com/1422-0067/24/8/7032 |
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