Oxidative stress induces mainly human centrin 2 polymerisation

Abstract

Purpose: To determine the human centrin 2 (Hscen 2) protein response to oxidising radicals in vitro and to evaluate the consequences on its biological functions.

Materials and methods: Hscen 2 was submitted to hydroxyl and azide radicals produced by radiolysis in the absence of oxygen. The resulting products were characterised by biochemical, spectroscopic and mass spectrometry techniques. Their thermodynamics parameters of complexation with C-terminal fragment of Xeroderma pigmentosum C protein (C-XPC), one of the Hscen 2 cellular partners, were quantified by isothermal titration calorimetry (ITC).

Results: Both hydroxyl and azide radicals induce centrin 2 polymerisation as we characterised several intermolecular cross-links generating dimers, trimers, tetramers and higher molecular mass species. These cross-links result from the formation of a covalent bond between the only tyrosine residue (Tyr 172) located in the C-terminal region of each monomer. Remarkably, dimerisation occurs for doses as low as a few grays. Moreover, this Hscen2 dimer has a lower affinity and stoechiometry binding to C-XPC.

Conclusions: These results show that as oxidative radicals induce high proportions of irreversible damages (polymerisation) centrin 2 is highly sensitive to ionising radiation. This could have important consequences on its biological functions.

Keywords:

• human centrin 2
• oxidative stress
• Xeroderma pigmentosum C
• bityrosine
• protein oxidation
• radiolysis

Acknowledgements

The authors thank C. Creascu for the access to circular dichroism and microcalorimetry apparatus.

Declaration of interest: The authors report no conflicts of interest. The authors alone are responsible for the content and writing of the paper.