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Light‐activated electron transfer and catalytic mechanism of carnitine oxidation by Rieske‐type oxygenase from human microbiota
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Shanmugam, Muralidharan, Quareshy, Mussa, Cameron, Alexander, Bugg, Timothy D. H. and Chen, Yin (2020) Light‐activated electron transfer and catalytic mechanism of carnitine oxidation by Rieske‐type oxygenase from human microbiota. Angewandte Chemie International Edition . doi:10.1002/anie.202012381 (In Press)
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WRAP-light‐activated-electron-transfer-catalytic-mechanism-carnitine-oxidation-Rieske‐type-oxygenase-human-microbiota-Chen-2020.pdf - Accepted Version Embargoed item. Restricted access to Repository staff only until 12 November 2021. Contact author directly, specifying your specific needs. - Requires a PDF viewer. Download (3025Kb) |
Official URL: https://doi.org/10.1002/anie.202012381
Abstract
Multi‐component, non‐heme iron‐dependent oxygenases catalyse a range of biochemical processes, as extensive as those found in heme chemistry. Oxidation of quaternary ammonium substrate carnitine by Acinetobacter baumannii (Ab) oxygenase CntA/ reductase CntB is implicated in the onset of human cardiovascular disease. However, detailed mechanistic understanding of the CntAB reaction is not known. Here, we develop a blue‐light (365 nm) activation of NADH coupled to electron paramagnetic resonance (EPR) measurements to study electron transfer from the excited state of NADH to the oxidised, Rieske‐type, [2Fe‐2S] 2+ cluster in the AbCntA oxygenase domain with and without the substrate, carnitine. Further electron transfer from one‐electron reduced, Rieske‐type [2Fe‐2S] 1+ centre in AbCntA‐WT to the mono‐nuclear, non‐heme iron centre via the bridging glutamate E205 and subsequent catalysis occurs only in the presence of carnitine. The reaction mechanism for photoactivated, NADH‐driven carnitine oxidation is different from the reaction driven by the AbCntB reductase partner. The electron transfer process in the AbCntA‐E205A mutant is severely affected, which plausibly accounts for the significant loss of catalytic activity in the AbCntA‐E205A mutant. The NADH photo‐activation coupled with EPR is broadly applicable to trap reactive intermediates at low temperature and creates a new method to characterise elusive intermediates in multiple redox‐centre containing proteins.
| Item Type: | Journal Article | ||||||
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| Subjects: | Q Science > QC Physics Q Science > QD Chemistry Q Science > QP Physiology |
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| Divisions: | Faculty of Science > Chemistry Faculty of Science > Life Sciences (2010- ) |
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| Library of Congress Subject Headings (LCSH): | Oxygenases, Metalloenzymes, Catalysis, Charge exchange, Carnitine | ||||||
| Journal or Publication Title: | Angewandte Chemie International Edition | ||||||
| Publisher: | Wiley - V C H Verlag GmbH & Co. KGaA | ||||||
| ISSN: | 1433-7851 | ||||||
| Official Date: | 12 November 2020 | ||||||
| Dates: |
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| Date of first compliant deposit: | 16 November 2020 | ||||||
| DOI: | 10.1002/anie.202012381 | ||||||
| Status: | Peer Reviewed | ||||||
| Publication Status: | In Press | ||||||
| Publisher Statement: | This is the peer reviewed version of the following article: Shanmugam, M., Quareshy, M., Cameron, A., Bugg, T..D.H. and Chen, Y. (2020), Light‐Activated Electron Transfer and Catalytic Mechanism of Carnitine Oxidation by Rieske‐type Oxygenase from Human Microbiota. Angew. Chem. Int. Ed.. Accepted Author Manuscript., which has been published in final form at https://doi.org/10.1002/anie.202012381. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived Versions. | ||||||
| Access rights to Published version: | Open Access | ||||||
| RIOXX Funder/Project Grant: |
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