Binding site for coenzyme A revealed in the structure of pyruvate:ferredoxin oxidoreductase from
Pyruvate:ferredoxin oxidoreductase (PFOR) is a microbial enzyme that uses thiamine pyrophosphate (TPP), three [4Fe-4S] clusters, and coenzyme A (CoA) in the reversible oxidation of pyruvate to generate acetyl-CoA and carbon dioxide. The two electrons that are generated as a result of pyruvate decarb...
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| Format: | Article |
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National Academy of Sciences (U.S.)
2018
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| Online Access: | http://hdl.handle.net/1721.1/119494 https://orcid.org/0000-0001-7362-9801 https://orcid.org/0000-0001-5486-2755 |
| _version_ | 1826202451961708544 |
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| author | Aman, Heather Can, Mehmet Ragsdale, Stephen W. Chen, Yang-Ting Drennan, Catherine L |
| author2 | Massachusetts Institute of Technology. Department of Biology |
| author_facet | Massachusetts Institute of Technology. Department of Biology Aman, Heather Can, Mehmet Ragsdale, Stephen W. Chen, Yang-Ting Drennan, Catherine L |
| author_sort | Aman, Heather |
| collection | MIT |
| description | Pyruvate:ferredoxin oxidoreductase (PFOR) is a microbial enzyme that uses thiamine pyrophosphate (TPP), three [4Fe-4S] clusters, and coenzyme A (CoA) in the reversible oxidation of pyruvate to generate acetyl-CoA and carbon dioxide. The two electrons that are generated as a result of pyruvate decarboxylation are used in the reduction of low potential ferredoxins, which provide reducing equivalents for central metabolism, including the Wood-Ljungdahl pathway. PFOR is a member of the 2-oxoacid:ferredoxin oxidoreductase (OFOR) superfamily, which plays major roles in both microbial redox reactions and carbon dioxide fixation. Here, we present a set of crystallographic snapshots of the best-studied member of this superfamily, the PFOR from Moorella thermoacetica (MtPFOR). These snapshots include the native structure, those of lactyl-TPP and acetyl-TPP reaction intermediates, and the first of an OFOR with CoA bound. These structural data reveal the binding site of CoA as domain III, the function of which in OFORs was previously unknown, and establish sequence motifs for CoA binding in the OFOR superfamily. MtPFOR structures further show that domain III undergoes a conformational change upon CoA binding that seals off the active site and positions the thiolate of CoA directly adjacent to the TPP cofactor. These structural findings provide a molecular basis for the experimental observation that CoA binding accelerates catalysis by 105-fold. Keywords: pyruvate:ferredoxin oxidoreductase; thiamine pyrophosphate; coenzyme A; gated electron transfer; carbon dioxide fixation |
| first_indexed | 2024-09-23T12:07:22Z |
| format | Article |
| id | mit-1721.1/119494 |
| institution | Massachusetts Institute of Technology |
| last_indexed | 2024-09-23T12:07:22Z |
| publishDate | 2018 |
| publisher | National Academy of Sciences (U.S.) |
| record_format | dspace |
| spelling | mit-1721.1/1194942022-10-01T08:22:27Z Binding site for coenzyme A revealed in the structure of pyruvate:ferredoxin oxidoreductase from Aman, Heather Can, Mehmet Ragsdale, Stephen W. Chen, Yang-Ting Drennan, Catherine L Massachusetts Institute of Technology. Department of Biology Massachusetts Institute of Technology. Department of Chemistry Chen, Yang-Ting Drennan, Catherine L Pyruvate:ferredoxin oxidoreductase (PFOR) is a microbial enzyme that uses thiamine pyrophosphate (TPP), three [4Fe-4S] clusters, and coenzyme A (CoA) in the reversible oxidation of pyruvate to generate acetyl-CoA and carbon dioxide. The two electrons that are generated as a result of pyruvate decarboxylation are used in the reduction of low potential ferredoxins, which provide reducing equivalents for central metabolism, including the Wood-Ljungdahl pathway. PFOR is a member of the 2-oxoacid:ferredoxin oxidoreductase (OFOR) superfamily, which plays major roles in both microbial redox reactions and carbon dioxide fixation. Here, we present a set of crystallographic snapshots of the best-studied member of this superfamily, the PFOR from Moorella thermoacetica (MtPFOR). These snapshots include the native structure, those of lactyl-TPP and acetyl-TPP reaction intermediates, and the first of an OFOR with CoA bound. These structural data reveal the binding site of CoA as domain III, the function of which in OFORs was previously unknown, and establish sequence motifs for CoA binding in the OFOR superfamily. MtPFOR structures further show that domain III undergoes a conformational change upon CoA binding that seals off the active site and positions the thiolate of CoA directly adjacent to the TPP cofactor. These structural findings provide a molecular basis for the experimental observation that CoA binding accelerates catalysis by 105-fold. Keywords: pyruvate:ferredoxin oxidoreductase; thiamine pyrophosphate; coenzyme A; gated electron transfer; carbon dioxide fixation National Institutes of Health (U.S.) (Grant GM069857) 2018-12-10T18:42:53Z 2018-12-10T18:42:53Z 2018-03 2018-12-04T15:43:01Z Article http://purl.org/eprint/type/JournalArticle 0027-8424 1091-6490 http://hdl.handle.net/1721.1/119494 Chen, Percival Yang-Ting et al. “Binding Site for Coenzyme A Revealed in the Structure of Pyruvate:ferredoxin Oxidoreductase from Moorella Thermoacetica.” Proceedings of the National Academy of Sciences 115,15 (March 2018): 3846–3851 © 2018 National Academy of Sciences https://orcid.org/0000-0001-7362-9801 https://orcid.org/0000-0001-5486-2755 http://dx.doi.org/10.1073/PNAS.1722329115 Proceedings of the National Academy of Sciences Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use. application/pdf National Academy of Sciences (U.S.) PNAS |
| spellingShingle | Aman, Heather Can, Mehmet Ragsdale, Stephen W. Chen, Yang-Ting Drennan, Catherine L Binding site for coenzyme A revealed in the structure of pyruvate:ferredoxin oxidoreductase from |
| title | Binding site for coenzyme A revealed in the structure of pyruvate:ferredoxin oxidoreductase from |
| title_full | Binding site for coenzyme A revealed in the structure of pyruvate:ferredoxin oxidoreductase from |
| title_fullStr | Binding site for coenzyme A revealed in the structure of pyruvate:ferredoxin oxidoreductase from |
| title_full_unstemmed | Binding site for coenzyme A revealed in the structure of pyruvate:ferredoxin oxidoreductase from |
| title_short | Binding site for coenzyme A revealed in the structure of pyruvate:ferredoxin oxidoreductase from |
| title_sort | binding site for coenzyme a revealed in the structure of pyruvate ferredoxin oxidoreductase from |
| url | http://hdl.handle.net/1721.1/119494 https://orcid.org/0000-0001-7362-9801 https://orcid.org/0000-0001-5486-2755 |
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