8R4H

Crystal structure of the copper efflux oxidase (CueO) from Hafnia alvei deleted of the Met-rich domain

Summary for 8R4H

• Entry DOI: 10.2210/pdb8r4h/pdb
• Related: 8R4F
• Descriptor: Copper efflux oxidase,Multicopper oxidase, COPPER (II) ION, 1,2-ETHANEDIOL, ... (4 entities in total)
• Functional Keywords: bacterial copper hemostasis, multi copper oxidase, metal binding protein
• Biological source: Hafnia alvei; More
• Total number of polymer chains: 2
• Total formula weight: 100941.85

Authors

Leone, P.,Contaldo, U. (deposition date: 2023-11-13, release date: 2025-04-23, Last modification date: 2025-11-05)

Primary citation

Contaldo, U.,Santucci, P.,Vergnes, A.,Leone, P.,Becam, J.,Biaso, F.,Ilbert, M.,Ezraty, B.,Lojou, E.,Mazurenko, I.
How the Larger Methionine-Rich Domain of CueO from Hafnia alvei Enhances Cuprous Oxidation.
Jacs Au, 5:1833-1844, 2025

PubMed Abstract: CueOs, members of the multicopper oxidase family, play a crucial role in bacterial copper detoxification. These enzymes feature a unique methionine-rich (Met-rich) domain, which is essential for the oxidation of Cu to Cu. Recent studies using CueO from (CueO) suggest that the Met-rich domain facilitates Cu recruitment from highly chelated species. To further explore this hypothesis, we produced and characterized a novel CueO from the bacterium (CueO). CueO possesses a significantly larger Met-rich domain than CueO, providing new insights into the role of this domain in cuprous oxidase activity. We first showed that CueO was as efficient in copper detoxification as CueO in vivo. The structures of both wild-type CueO and a variant lacking the Met-rich domain were resolved by X-ray crystallography and simulated by molecular dynamics, offering a detailed structural basis for understanding their functions. Cuprous oxidase activity was then quantified either from free electrogenerated Cu with CueO immobilized on an electrode or from different Cu-complexes with CueO in solution. These methods enabled the fine-tuning of Cu chelation strength. Consistent with findings for CueO, it was confirmed that the Met-rich domain of CueO is dispensable for Cu oxidation when weakly chelated Cu is used. However, its role becomes crucial as chelation strength increases. Comparative analyses of cuprous oxidase activity between CueO and CueO revealed that CueO outperforms CueO, demonstrating superior efficiency in oxidizing Cu from chelated forms. This enhanced activity correlates with the higher methionine content in CueO, which appears to play a pivotal role in facilitating Cu oxidation under conditions of stronger chelation.

PubMed: 40313819
DOI: 10.1021/jacsau.5c00076

Experimental method

X-RAY DIFFRACTION (2.937 Å)