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Serial femtosecond zero dose crystallography captures a water‐free distal heme site in a dye‐decolorising peroxidase to reveal a catalytic role for an arginine in FeIV=O Formation

Serial femtosecond zero dose crystallography captures a water‐free distal heme site in a dye‐decolorising peroxidase to reveal a catalytic role for an arginine in FeIV=O Formation
Serial femtosecond zero dose crystallography captures a water‐free distal heme site in a dye‐decolorising peroxidase to reveal a catalytic role for an arginine in FeIV=O Formation
Obtaining structures of intact redox states of metal centers derived from zero dose X‐ray crystallography can advance our mechanistic understanding of metalloenzymes. In dye‐decolorising heme peroxidases (DyPs), controversy exists regarding the mechanistic role of the distal heme residues aspartate and arginine in the heterolysis of peroxide to form the catalytic intermediate compound I (FeIV=O and a porphyrin cation radical). Using serial femtosecond X‐ray crystallography (SFX), we have determined the pristine structures of the FeIII and FeIV=O redox states of a B‐type DyP. These structures reveal a water‐free distal heme site that, together with the presence of an asparagine, imply the use of the distal arginine as a catalytic base. A combination of mutagenesis and kinetic studies corroborate such a role. Our SFX approach thus provides unique insight into how the distal heme site of DyPs can be tuned to select aspartate or arginine for the rate enhancement of peroxide heterolysis.
1433-7851
21656–21662
Lučić, Marina
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Svistunenko, Dimitri A.
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Wilson, Michael T.
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Chaplin, Amanda K.
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Davy, Bradley
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Ebrahim, Ali
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Axford, Danny
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Tosha, Takehiko
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Sugimoto, Hiroshi
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Owada, Shigeki
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Dworkowski, Florian S. N
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Tews, Ivo
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Owen, Robin L.
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Hough, Michael A.
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Worrall, Jonathan A. R.
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Lučić, Marina
0a60f017-9f9c-4a1c-bff7-d8b062fdd4e1
Svistunenko, Dimitri A.
fe718070-aa4f-48d4-b951-7e86c5478304
Wilson, Michael T.
159ea454-389c-460f-bd5d-54fcddb448e2
Chaplin, Amanda K.
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Davy, Bradley
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Ebrahim, Ali
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Axford, Danny
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Tosha, Takehiko
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Sugimoto, Hiroshi
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Owada, Shigeki
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Dworkowski, Florian S. N
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Tews, Ivo
9117fc5e-d01c-4f8d-a734-5b14d3eee8dd
Owen, Robin L.
101c8487-b6fb-4930-a223-8c569db19536
Hough, Michael A.
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Worrall, Jonathan A. R.
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Lučić, Marina, Svistunenko, Dimitri A., Wilson, Michael T., Chaplin, Amanda K., Davy, Bradley, Ebrahim, Ali, Axford, Danny, Tosha, Takehiko, Sugimoto, Hiroshi, Owada, Shigeki, Dworkowski, Florian S. N, Tews, Ivo, Owen, Robin L., Hough, Michael A. and Worrall, Jonathan A. R. (2020) Serial femtosecond zero dose crystallography captures a water‐free distal heme site in a dye‐decolorising peroxidase to reveal a catalytic role for an arginine in FeIV=O Formation. Angewandte Chemie International Edition, 59 (48), 21656–21662. (doi:10.1002/anie.202008622).

Record type: Article

Abstract

Obtaining structures of intact redox states of metal centers derived from zero dose X‐ray crystallography can advance our mechanistic understanding of metalloenzymes. In dye‐decolorising heme peroxidases (DyPs), controversy exists regarding the mechanistic role of the distal heme residues aspartate and arginine in the heterolysis of peroxide to form the catalytic intermediate compound I (FeIV=O and a porphyrin cation radical). Using serial femtosecond X‐ray crystallography (SFX), we have determined the pristine structures of the FeIII and FeIV=O redox states of a B‐type DyP. These structures reveal a water‐free distal heme site that, together with the presence of an asparagine, imply the use of the distal arginine as a catalytic base. A combination of mutagenesis and kinetic studies corroborate such a role. Our SFX approach thus provides unique insight into how the distal heme site of DyPs can be tuned to select aspartate or arginine for the rate enhancement of peroxide heterolysis.

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More information

Published date: 11 August 2020

Identifiers

Local EPrints ID: 447414
URI: http://eprints.soton.ac.uk/id/eprint/447414
ISSN: 1433-7851
PURE UUID: b3276237-bc8d-4a16-a503-9ca7cb6488fc
ORCID for Ivo Tews: ORCID iD orcid.org/0000-0002-4704-1139

Catalogue record

Date deposited: 11 Mar 2021 17:30
Last modified: 26 Nov 2021 02:56

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Contributors

Author: Marina Lučić
Author: Dimitri A. Svistunenko
Author: Michael T. Wilson
Author: Amanda K. Chaplin
Author: Bradley Davy
Author: Ali Ebrahim
Author: Danny Axford
Author: Takehiko Tosha
Author: Hiroshi Sugimoto
Author: Shigeki Owada
Author: Florian S. N Dworkowski
Author: Ivo Tews ORCID iD
Author: Robin L. Owen
Author: Michael A. Hough
Author: Jonathan A. R. Worrall

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