Postbiosynthetic modification of a precursor to the nitrogenase iron–molybdenum cofactor
© 2021 National Academy of Sciences. All rights reserved. Nitrogenases utilize Fe-S clusters to reduce N2to NH3. The large number of Fe sites in their catalytic cofactors has hampered spectroscopic investigations into their electronic structures, mechanisms, and biosyntheses. To facilitate their spe...
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| Format: | Article |
| Language: | English |
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Proceedings of the National Academy of Sciences
2022
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| Online Access: | https://hdl.handle.net/1721.1/141323 |
| _version_ | 1811089286103040000 |
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| author | Srisantitham, Suppachai Badding, Edward D Suess, Daniel LM |
| author2 | Massachusetts Institute of Technology. Department of Chemistry |
| author_facet | Massachusetts Institute of Technology. Department of Chemistry Srisantitham, Suppachai Badding, Edward D Suess, Daniel LM |
| author_sort | Srisantitham, Suppachai |
| collection | MIT |
| description | © 2021 National Academy of Sciences. All rights reserved. Nitrogenases utilize Fe-S clusters to reduce N2to NH3. The large number of Fe sites in their catalytic cofactors has hampered spectroscopic investigations into their electronic structures, mechanisms, and biosyntheses. To facilitate their spectroscopic analysis, we are developing methods for incorporating 57Fe into specific sites of nitrogenase cofactors, and we report herein siteselective 57Fe labeling of the L-cluster-a carbide-containing, [Fe8S9C] precursor to the Mo nitrogenase catalytic cofactor. Treatment of the isolated L-cluster with the chelator ethylenediaminetetraacetate followed by reconstitution with 57Fe2+results in 57Fe labeling of the terminal Fe sites in high yield and with high selectivity. This protocol enables the generation of L-cluster samples in which either the two terminal or the six belt Fe sites are selectively labeled with 57Fe. Mössbauer spectroscopic analysis of these samples bound to the nitrogenase maturase Azotobacter vinelandii NifX reveals differences in the primary coordination sphere of the terminal Fe sites and that one of the terminal sites of the L-cluster binds to H35 of Av NifX. This work provides molecularlevel insights into the electronic structure and biosynthesis of the L-cluster and introduces postbiosynthetic modification as a promising strategy for studies of nitrogenase cofactors. |
| first_indexed | 2024-09-23T14:16:47Z |
| format | Article |
| id | mit-1721.1/141323 |
| institution | Massachusetts Institute of Technology |
| language | English |
| last_indexed | 2024-09-23T14:16:47Z |
| publishDate | 2022 |
| publisher | Proceedings of the National Academy of Sciences |
| record_format | dspace |
| spelling | mit-1721.1/1413232023-02-10T20:34:34Z Postbiosynthetic modification of a precursor to the nitrogenase iron–molybdenum cofactor Srisantitham, Suppachai Badding, Edward D Suess, Daniel LM Massachusetts Institute of Technology. Department of Chemistry © 2021 National Academy of Sciences. All rights reserved. Nitrogenases utilize Fe-S clusters to reduce N2to NH3. The large number of Fe sites in their catalytic cofactors has hampered spectroscopic investigations into their electronic structures, mechanisms, and biosyntheses. To facilitate their spectroscopic analysis, we are developing methods for incorporating 57Fe into specific sites of nitrogenase cofactors, and we report herein siteselective 57Fe labeling of the L-cluster-a carbide-containing, [Fe8S9C] precursor to the Mo nitrogenase catalytic cofactor. Treatment of the isolated L-cluster with the chelator ethylenediaminetetraacetate followed by reconstitution with 57Fe2+results in 57Fe labeling of the terminal Fe sites in high yield and with high selectivity. This protocol enables the generation of L-cluster samples in which either the two terminal or the six belt Fe sites are selectively labeled with 57Fe. Mössbauer spectroscopic analysis of these samples bound to the nitrogenase maturase Azotobacter vinelandii NifX reveals differences in the primary coordination sphere of the terminal Fe sites and that one of the terminal sites of the L-cluster binds to H35 of Av NifX. This work provides molecularlevel insights into the electronic structure and biosynthesis of the L-cluster and introduces postbiosynthetic modification as a promising strategy for studies of nitrogenase cofactors. 2022-03-21T14:50:11Z 2022-03-21T14:50:11Z 2021 2022-03-21T14:39:34Z Article http://purl.org/eprint/type/JournalArticle https://hdl.handle.net/1721.1/141323 Srisantitham, Suppachai, Badding, Edward D and Suess, Daniel LM. 2021. "Postbiosynthetic modification of a precursor to the nitrogenase iron–molybdenum cofactor." Proceedings of the National Academy of Sciences of the United States of America, 118 (11). en 10.1073/PNAS.2015361118 Proceedings of the National Academy of Sciences of the United States of America 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 Proceedings of the National Academy of Sciences PNAS |
| spellingShingle | Srisantitham, Suppachai Badding, Edward D Suess, Daniel LM Postbiosynthetic modification of a precursor to the nitrogenase iron–molybdenum cofactor |
| title | Postbiosynthetic modification of a precursor to the nitrogenase iron–molybdenum cofactor |
| title_full | Postbiosynthetic modification of a precursor to the nitrogenase iron–molybdenum cofactor |
| title_fullStr | Postbiosynthetic modification of a precursor to the nitrogenase iron–molybdenum cofactor |
| title_full_unstemmed | Postbiosynthetic modification of a precursor to the nitrogenase iron–molybdenum cofactor |
| title_short | Postbiosynthetic modification of a precursor to the nitrogenase iron–molybdenum cofactor |
| title_sort | postbiosynthetic modification of a precursor to the nitrogenase iron molybdenum cofactor |
| url | https://hdl.handle.net/1721.1/141323 |
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