Unraveling the impact of cysteine-to-serine mutations on the structural and functional properties of Cu(I)-binding proteins

Matic Pavlin, Zena Qasem, Hila Sameach, Lada Gevorkyan-Airapetov, Ida Ritacco, Sharon Ruthstein, Alessandra Magistrato

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8 Scopus citations


Appropriate maintenance of Cu(I) homeostasis is an essential requirement for proper cell function because its misregulation induces the onset of major human diseases and mortality. For this reason, several research efforts have been devoted to dissecting the inner working mechanism of Cu(I)-binding proteins and transporters. A commonly adopted strategy relies on mutations of cysteine residues, for which Cu(I) has an exquisite complementarity, to serines. Nevertheless, in spite of the similarity between these two amino acids, the structural and functional impact of serine mutations on Cu(I)-binding biomolecules remains unclear. Here, we applied various biochemical and biophysical methods, together with all-atom simulations, to investigate the effect of these mutations on the stability, structure, and aggregation propensity of Cu(I)-binding proteins, as well as their interaction with specific partner proteins. Among Cu(I)-binding biomolecules, we focused on the eukaryotic Atox1-ATP7B system, and the prokaryotic CueR metalloregulator. Our results reveal that proteins containing cysteine-to-serine mutations can still bind Cu(I) ions; however, this alters their stability and aggregation propensity. These results contribute to deciphering the critical biological principles underlying the regulatory mechanism of the in-cell Cu(I) concentration, and provide a basis for interpreting future studies that will take advantage of cysteine-to-serine mutations in Cu(I)-binding systems.

Original languageEnglish
Article number3462
JournalInternational Journal of Molecular Sciences
Issue number14
StatePublished - 2 Jul 2019

Bibliographical note

Funding Information:
Funding: A.M. and S.R. thank the Italian Ministry for International affairs and Israel Ministry of Science for the project Cu(Metabolism). A.M. thanks the Italian Association for Cancer Research (my First AIRC grant no 17134). This research was funded by the ERC-STG no. 754365 given to S.R.

Publisher Copyright:
© 2019 by the authors. Licensee MDPI, Basel, Switzerland.


  • ATP7B
  • Atox1
  • Copper metabolism
  • Cu(I) affinity
  • CueR
  • EPR
  • QM/MM
  • Serine mutations
  • molecular dynamics


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