Design principles for site-selective hydroxylation by a Rieske oxygenase.


Journal

Nature communications
ISSN: 2041-1723
Titre abrégé: Nat Commun
Pays: England
ID NLM: 101528555

Informations de publication

Date de publication:
11 01 2022
Historique:
received: 07 08 2021
accepted: 15 12 2021
entrez: 12 1 2022
pubmed: 13 1 2022
medline: 11 2 2022
Statut: epublish

Résumé

Rieske oxygenases exploit the reactivity of iron to perform chemically challenging C-H bond functionalization reactions. Thus far, only a handful of Rieske oxygenases have been structurally characterized and remarkably little information exists regarding how these enzymes use a common architecture and set of metallocenters to facilitate a diverse range of reactions. Herein, we detail how two Rieske oxygenases SxtT and GxtA use different protein regions to influence the site-selectivity of their catalyzed monohydroxylation reactions. We present high resolution crystal structures of SxtT and GxtA with the native β-saxitoxinol and saxitoxin substrates bound in addition to a Xenon-pressurized structure of GxtA that reveals the location of a substrate access tunnel to the active site. Ultimately, this structural information allowed for the identification of six residues distributed between three regions of SxtT that together control the selectivity of the C-H hydroxylation event. Substitution of these residues produces a SxtT variant that is fully adapted to exhibit the non-native site-selectivity and substrate scope of GxtA. Importantly, we also found that these selectivity regions are conserved in other structurally characterized Rieske oxygenases, providing a framework for predictively repurposing and manipulating Rieske oxygenases as biocatalysts.

Identifiants

pubmed: 35017498
doi: 10.1038/s41467-021-27822-3
pii: 10.1038/s41467-021-27822-3
pmc: PMC8752792
doi:

Substances chimiques

Escherichia coli Proteins 0
Sxy protein, E coli 0
Trans-Activators 0
Iron E1UOL152H7
Oxygenases EC 1.13.-

Types de publication

Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.

Langues

eng

Sous-ensembles de citation

IM

Pagination

255

Subventions

Organisme : NINDS NIH HHS
ID : F31 NS111906
Pays : United States
Organisme : NIGMS NIH HHS
ID : P30 GM133894
Pays : United States
Organisme : NIGMS NIH HHS
ID : R35 GM124880
Pays : United States
Organisme : NIGMS NIH HHS
ID : R35 GM138271
Pays : United States

Informations de copyright

© 2022. The Author(s).

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Auteurs

Jianxin Liu (J)

Department of Chemistry, University of Michigan, Ann Arbor, MI, 48109, USA.

Jiayi Tian (J)

Department of Chemistry, University of Michigan, Ann Arbor, MI, 48109, USA.

Christopher Perry (C)

Department of Chemistry, University of Michigan, Ann Arbor, MI, 48109, USA.
Program in Chemical Biology, University of Michigan, Ann Arbor, MI, 48109, USA.
Life Sciences Institute, University of Michigan, Ann Arbor, MI, 48109, USA.

April L Lukowski (AL)

Program in Chemical Biology, University of Michigan, Ann Arbor, MI, 48109, USA.
Life Sciences Institute, University of Michigan, Ann Arbor, MI, 48109, USA.
Center for Marine Biotechnology and Biomedicine, Scripps Institution of Oceanography, University of California, San Diego, La Jolla, CA, 92037, USA.

Tzanko I Doukov (TI)

Macromolecular Crystallography Group, Stanford Synchrotron Radiation Light Source, SLAC National Accelerator Laboratory, Menlo Park, CA, 94025, USA.

Alison R H Narayan (ARH)

Department of Chemistry, University of Michigan, Ann Arbor, MI, 48109, USA. arhardin@umich.edu.
Program in Chemical Biology, University of Michigan, Ann Arbor, MI, 48109, USA. arhardin@umich.edu.
Life Sciences Institute, University of Michigan, Ann Arbor, MI, 48109, USA. arhardin@umich.edu.

Jennifer Bridwell-Rabb (J)

Department of Chemistry, University of Michigan, Ann Arbor, MI, 48109, USA. jebridwe@umich.edu.
Program in Chemical Biology, University of Michigan, Ann Arbor, MI, 48109, USA. jebridwe@umich.edu.
Life Sciences Institute, University of Michigan, Ann Arbor, MI, 48109, USA. jebridwe@umich.edu.

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