Electrochemical Characterization of Escherichia coli Adaptive Response Protein AidB
When exposed to known DNA-damaging alkylating agents, Escherichia coli cells increase production of four DNA repair enzymes: Ada, AlkA, AlkB, and AidB. The role of three enzymes (Ada, AlkA, and AlkB) in repairing DNA lesions has been well characterized, while the function of AidB is poorly understoo...
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| Format: | Article |
| Language: | en_US |
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MDPI AG
2013
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| Online Access: | http://hdl.handle.net/1721.1/77576 https://orcid.org/0000-0001-5486-2755 |
| _version_ | 1826214979259334656 |
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| author | Hamill, Michael J. Jost, Marco Wong, Cintyu Bene, Nicholas C. Elliott, Sean J. Drennan, Catherine L |
| author2 | Massachusetts Institute of Technology. Center for Environmental Health Sciences |
| author_facet | Massachusetts Institute of Technology. Center for Environmental Health Sciences Hamill, Michael J. Jost, Marco Wong, Cintyu Bene, Nicholas C. Elliott, Sean J. Drennan, Catherine L |
| author_sort | Hamill, Michael J. |
| collection | MIT |
| description | When exposed to known DNA-damaging alkylating agents, Escherichia coli cells increase production of four DNA repair enzymes: Ada, AlkA, AlkB, and AidB. The role of three enzymes (Ada, AlkA, and AlkB) in repairing DNA lesions has been well characterized, while the function of AidB is poorly understood. AidB has a distinct cofactor that is potentially related to the elusive role of AidB in adaptive response: a redox active flavin adenine dinucleotide (FAD). In this study, we report the thermodynamic redox properties of the AidB flavin for the first time, both for free protein and in the presence of potential substrates. We find that the midpoint reduction potential of the AidB flavin is within a biologically relevant window for redox chemistry at −181 mV, that AidB significantly stabilizes the flavin semiquinone, and that small molecule binding perturbs the observed reduction potential. Our electrochemical results combined with structural analysis allow for fresh comparisons between AidB and the homologous acyl-coenzyme A dehydrogenase (ACAD) family of enzymes. AidB exhibits several discrepancies from ACADs that suggest a novel catalytic mechanism distinct from that of the ACAD family enzymes. |
| first_indexed | 2024-09-23T16:14:20Z |
| format | Article |
| id | mit-1721.1/77576 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T16:14:20Z |
| publishDate | 2013 |
| publisher | MDPI AG |
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| spelling | mit-1721.1/775762022-10-02T07:11:15Z Electrochemical Characterization of Escherichia coli Adaptive Response Protein AidB Hamill, Michael J. Jost, Marco Wong, Cintyu Bene, Nicholas C. Elliott, Sean J. Drennan, Catherine L Massachusetts Institute of Technology. Center for Environmental Health Sciences Massachusetts Institute of Technology. Department of Biology Massachusetts Institute of Technology. Department of Chemistry Massachusetts Institute of Technology. School of Science Hamill, Michael J. Jost, Marco Wong, Cintyu Drennan, Catherine L. When exposed to known DNA-damaging alkylating agents, Escherichia coli cells increase production of four DNA repair enzymes: Ada, AlkA, AlkB, and AidB. The role of three enzymes (Ada, AlkA, and AlkB) in repairing DNA lesions has been well characterized, while the function of AidB is poorly understood. AidB has a distinct cofactor that is potentially related to the elusive role of AidB in adaptive response: a redox active flavin adenine dinucleotide (FAD). In this study, we report the thermodynamic redox properties of the AidB flavin for the first time, both for free protein and in the presence of potential substrates. We find that the midpoint reduction potential of the AidB flavin is within a biologically relevant window for redox chemistry at −181 mV, that AidB significantly stabilizes the flavin semiquinone, and that small molecule binding perturbs the observed reduction potential. Our electrochemical results combined with structural analysis allow for fresh comparisons between AidB and the homologous acyl-coenzyme A dehydrogenase (ACAD) family of enzymes. AidB exhibits several discrepancies from ACADs that suggest a novel catalytic mechanism distinct from that of the ACAD family enzymes. National Institutes of Health (U.S.) (Grant P30-ES002109) National Institutes of Health (U.S.) (Grant R01-GM69857) National Science Foundation (U.S.) (Grant MCB-0543833) 2013-03-06T16:50:49Z 2013-03-06T16:50:49Z 2012-12 2012-11 Article http://purl.org/eprint/type/JournalArticle 1422-0067 http://hdl.handle.net/1721.1/77576 Hamill, Michael et al. “Electrochemical Characterization of Escherichia Coli Adaptive Response Protein AidB.” International Journal of Molecular Sciences 13.12 (2012): 16899–16915. https://orcid.org/0000-0001-5486-2755 en_US http://dx.doi.org/10.3390/ijms131216899 International Journal of Molecular Sciences Creative Commons Attribution 3.0 http://creativecommons.org/licenses/by/3.0/ application/pdf MDPI AG MDPI Publishing |
| spellingShingle | Hamill, Michael J. Jost, Marco Wong, Cintyu Bene, Nicholas C. Elliott, Sean J. Drennan, Catherine L Electrochemical Characterization of Escherichia coli Adaptive Response Protein AidB |
| title | Electrochemical Characterization of Escherichia coli Adaptive Response Protein AidB |
| title_full | Electrochemical Characterization of Escherichia coli Adaptive Response Protein AidB |
| title_fullStr | Electrochemical Characterization of Escherichia coli Adaptive Response Protein AidB |
| title_full_unstemmed | Electrochemical Characterization of Escherichia coli Adaptive Response Protein AidB |
| title_short | Electrochemical Characterization of Escherichia coli Adaptive Response Protein AidB |
| title_sort | electrochemical characterization of escherichia coli adaptive response protein aidb |
| url | http://hdl.handle.net/1721.1/77576 https://orcid.org/0000-0001-5486-2755 |
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