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Crystal structure of ferric recombinant horseradish peroxidase

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posted on 2025-04-02, 08:25 authored by Mst Luthfun Nesa, Suman K Mandal, Christine Toelzer, Diana Humer, Peter MoodyPeter Moody, Imre Berger, Oliver Spadiut, Emma L Raven

Horseradish peroxidase (HRP), isolated from horseradish roots, is heavily glycosylated, making it difficult to crystallize. In this work, we produced recombinant HRP in E. coli and obtained an X-ray structure of the ferric enzyme at 1.63 Å resolution. The structure shows that the recombinant HRP contains four disulphide bonds and two calcium ions, which are highly conserved in class III peroxidase enzymes. The heme active site contains histidine residues at the proximal (His 170) and distal (His 42) positions, and an active site arginine (Arg 38). Surprisingly, an ethylene glycol molecule was identified in the active site, forming hydrogen bonds with His 42 and Arg 38 at the δ-heme edge. The high yields obtained from the recombinant expression system, and the successful crystallization of the enzyme pave the way for new structural studies in the future. Graphical abstract

Funding

Leverhulme Trust (grant 2024-263/4 to ER)

PhD Scholarship from the University of Bristol

History

Author affiliation

College of Life Sciences Molecular & Cell Biology

Version

  • VoR (Version of Record)

Published in

JBIC Journal of Biological Inorganic Chemistry

Publisher

Springer Science and Business Media LLC

issn

0949-8257

eissn

1432-1327

Copyright date

2025

Available date

2025-04-02

Spatial coverage

Germany

Language

en

Deposited by

Professor Peter Moody

Deposit date

2025-03-27

Data Access Statement

The crystallographic data for the structures discussed in this article have been deposited at the wwPDB protein data bank under the deposition PDB ID 9H1M. The X-ray structure validation report has been submitted in the related files section. All other relevant data generated and analyzed during this study, including experimental, spectroscopic, and crystallographic data, are included in this article and its supplementary information.

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