Publication

Article Metrics

Citations


Online attention

Relations between structure and Zn(II) binding affinity shed light on the mechanisms of Rad50 hook domain functioning and its phosphorylation

DOI: 10.3390/ijms231911140 DOI Help

Authors: Jozef Ba Tran (Univeristy of Wroclaw) , Michal Padjasek (Univeristy of Wroclaw) , Artur Krezel (Univeristy of Wroclaw)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: International Journal Of Molecular Sciences , VOL 23

State: Published (Approved)
Published: October 2022
Diamond Proposal Number(s): 22570

Open Access Open Access

Abstract: The metal binding at protein–protein interfaces is still uncharted territory in intermolecular interactions. To date, only a few protein complexes binding Zn(II) in an intermolecular manner have been deeply investigated. The most notable example of such interfaces is located in the highly conserved Rad50 protein, part of the Mre11-Rad50-Nbs1 (MRN) complex, where Zn(II) is required for homodimerization (Zn(Rad50)2). The high stability of Zn(Rad50)2 is conserved not only for the protein derived from the thermophilic archaeon Pyrococcus furiosus (logK12 = 20.95 for 130-amino-acid-long fragment), which was the first one studied, but also for the human paralog studied here (logK12 = 19.52 for a 183-amino-acid-long fragment). As we reported previously, the extremely high stability results from the metal-coupled folding process where particular Rad50 protein fragments play a critical role. The sequence–structure–stability analysis based on human Rad50 presented here separates the individual structural components that increase the stability of the complex, pointing to amino acid residues far away from the Zn(II) binding site as being largely responsible for the complex stabilization. The influence of the individual components is very well reflected by the previously published crystal structure of the human Rad50 zinc hook (PDB: 5GOX). In addition, we hereby report the effect of phosphorylation of the zinc hook domain, which exerts a destabilizing effect on the domain. This study identifies factors governing the stability of metal-mediated protein–protein interactions and illuminates their molecular basis.

Journal Keywords: DNA damage; Rad50; zinc; zinc hook; Mre11-Rad50-Nbs1 (MRN)

Subject Areas: Biology and Bio-materials, Chemistry


Instruments: B18-Core EXAFS , B21-High Throughput SAXS

Added On: 29/09/2022 11:43

Documents:
ijms-23-11140.pdf

Discipline Tags:

Biochemistry Chemistry Biophysics Life Sciences & Biotech

Technical Tags:

Spectroscopy X-ray Absorption Spectroscopy (XAS) Extended X-ray Absorption Fine Structure (EXAFS) X-ray Absorption Near Edge Structure (XANES)