Gated Proton Release during Radical Transfer at the Subunit Interface of Ribonucleotide Reductase
© The class Ia ribonucleotide reductase of Escherichia coli requires strict regulation of long-range radical transfer between two subunits, α and β, through a series of redox-active amino acids (Y122•[β] ↔ W48?[β] ↔ Y356[β] ↔ Y731[α] ↔ Y730[α] ↔ C439[α]). Nowhere is this more precarious than at the...
| Príomhchruthaitheoirí: | , , , , , |
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| Formáid: | Alt |
| Teanga: | English |
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American Chemical Society (ACS)
2022
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| Rochtain ar líne: | https://hdl.handle.net/1721.1/146788 |
| _version_ | 1826210297804750848 |
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| author | Cui, Chang Greene, Brandon L Kang, Gyunghoon Drennan, Catherine L Stubbe, JoAnne Nocera, Daniel G |
| author2 | Massachusetts Institute of Technology. Department of Biology |
| author_facet | Massachusetts Institute of Technology. Department of Biology Cui, Chang Greene, Brandon L Kang, Gyunghoon Drennan, Catherine L Stubbe, JoAnne Nocera, Daniel G |
| author_sort | Cui, Chang |
| collection | MIT |
| description | © The class Ia ribonucleotide reductase of Escherichia coli requires strict regulation of long-range radical transfer between two subunits, α and β, through a series of redox-active amino acids (Y122•[β] ↔ W48?[β] ↔ Y356[β] ↔ Y731[α] ↔ Y730[α] ↔ C439[α]). Nowhere is this more precarious than at the subunit interface. Here, we show that the oxidation of Y356 is regulated by proton release involving a specific residue, E52[β], which is part of a water channel at the subunit interface for rapid proton transfer to the bulk solvent. An E52Q variant is incapable of Y356 oxidation via the native radical transfer pathway or non-native photochemical oxidation, following photosensitization by covalent attachment of a photo-oxidant at position 355[β]. Substitution of Y356 for various FnY analogues in an E52Q-photoβ2, where the side chain remains deprotonated, recovered photochemical enzymatic turnover. Transient absorption and emission data support the conclusion that Y356 oxidation requires E52 for proton management, suggesting its essential role in gating radical transport across the protein-protein interface. |
| first_indexed | 2024-09-23T14:47:34Z |
| format | Article |
| id | mit-1721.1/146788 |
| institution | Massachusetts Institute of Technology |
| language | English |
| last_indexed | 2024-09-23T14:47:34Z |
| publishDate | 2022 |
| publisher | American Chemical Society (ACS) |
| record_format | dspace |
| spelling | mit-1721.1/1467882022-12-08T03:11:44Z Gated Proton Release during Radical Transfer at the Subunit Interface of Ribonucleotide Reductase Cui, Chang Greene, Brandon L Kang, Gyunghoon Drennan, Catherine L Stubbe, JoAnne Nocera, Daniel G Massachusetts Institute of Technology. Department of Biology Massachusetts Institute of Technology. Department of Chemistry © The class Ia ribonucleotide reductase of Escherichia coli requires strict regulation of long-range radical transfer between two subunits, α and β, through a series of redox-active amino acids (Y122•[β] ↔ W48?[β] ↔ Y356[β] ↔ Y731[α] ↔ Y730[α] ↔ C439[α]). Nowhere is this more precarious than at the subunit interface. Here, we show that the oxidation of Y356 is regulated by proton release involving a specific residue, E52[β], which is part of a water channel at the subunit interface for rapid proton transfer to the bulk solvent. An E52Q variant is incapable of Y356 oxidation via the native radical transfer pathway or non-native photochemical oxidation, following photosensitization by covalent attachment of a photo-oxidant at position 355[β]. Substitution of Y356 for various FnY analogues in an E52Q-photoβ2, where the side chain remains deprotonated, recovered photochemical enzymatic turnover. Transient absorption and emission data support the conclusion that Y356 oxidation requires E52 for proton management, suggesting its essential role in gating radical transport across the protein-protein interface. 2022-12-07T18:23:54Z 2022-12-07T18:23:54Z 2021 2022-12-07T18:17:10Z Article http://purl.org/eprint/type/JournalArticle https://hdl.handle.net/1721.1/146788 Cui, Chang, Greene, Brandon L, Kang, Gyunghoon, Drennan, Catherine L, Stubbe, JoAnne et al. 2021. "Gated Proton Release during Radical Transfer at the Subunit Interface of Ribonucleotide Reductase." Journal of the American Chemical Society, 143 (1). en 10.1021/JACS.0C07879 Journal of the American Chemical Society Creative Commons Attribution 4.0 International license https://creativecommons.org/licenses/by/4.0/ application/pdf American Chemical Society (ACS) PMC |
| spellingShingle | Cui, Chang Greene, Brandon L Kang, Gyunghoon Drennan, Catherine L Stubbe, JoAnne Nocera, Daniel G Gated Proton Release during Radical Transfer at the Subunit Interface of Ribonucleotide Reductase |
| title | Gated Proton Release during Radical Transfer at the Subunit Interface of Ribonucleotide Reductase |
| title_full | Gated Proton Release during Radical Transfer at the Subunit Interface of Ribonucleotide Reductase |
| title_fullStr | Gated Proton Release during Radical Transfer at the Subunit Interface of Ribonucleotide Reductase |
| title_full_unstemmed | Gated Proton Release during Radical Transfer at the Subunit Interface of Ribonucleotide Reductase |
| title_short | Gated Proton Release during Radical Transfer at the Subunit Interface of Ribonucleotide Reductase |
| title_sort | gated proton release during radical transfer at the subunit interface of ribonucleotide reductase |
| url | https://hdl.handle.net/1721.1/146788 |
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