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The C-Terminus of Human Copper Importer Ctr1 Acts as a Binding Site and Transfers Copper to Atox1
Umeå University, Faculty of Science and Technology, Department of Chemistry.
Umeå University, Faculty of Science and Technology, Department of Chemistry. Chemistry Department, University of Konstanz, Konstanz, Germany.
Umeå University, Faculty of Science and Technology, Department of Chemistry. Department of Biology and Biological Engineering, Chalmers University of Technology, Gothenburg, Sweden.
2016 (English)In: Biophysical Journal, ISSN 0006-3495, E-ISSN 1542-0086, Vol. 110, no 1, 95-102 p.Article in journal (Refereed) PublishedText
Abstract [en]

Uptake of copper (Cu) ions into human cells is mediated by the plasma membrane protein Ctr1 and is followed by Cu transfer to cytoplasmic Cu chaperones for delivery to Cu-dependent enzymes. The C-terminal cytoplasmic tail of Ctr1 is a 13-residue peptide harboring an HCH motif that is thought to interact with Cu. We here employ biophysical experiments under anaerobic conditions in peptide models of the Ctr1 C-terminus to deduce Cu-binding residues, Cu affinity, and the ability to release Cu to the cytoplasmic Cu chaperone Atox1. Based on NMR assignments and bicinchoninic acid competition experiments, we demonstrate that Cu interacts in a 1:1 stoichiometry with the HCH motif with an affinity, K-D, of similar to 10(-14) M. Removing either the Cys residue or the two His residues lowers the Cu-peptide affinity, but site specificity is retained. The C-terminal peptide and Atox1 do not interact in solution in the absence of Cu. However, as directly demonstrated at the residue level via NMR spectroscopy, Atox1 readily acquires Cu from the Cu-loaded peptide. We propose that Cu binding to the Ctr1 C-terminal tail regulates Cu transport into the cytoplasm such that the metal ion is only released to high-affinity Cu chaperones.

Place, publisher, year, edition, pages
2016. Vol. 110, no 1, 95-102 p.
National Category
Biophysics
Identifiers
URN: urn:nbn:se:umu:diva-114887DOI: 10.1016/j.bpj.2015.11.016ISI: 000367783900032PubMedID: 26745413OAI: oai:DiVA.org:umu-114887DiVA: diva2:923798
Available from: 2016-04-27 Created: 2016-01-29 Last updated: 2016-04-27Bibliographically approved

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Kahra, DanaKovermann, MichaelWittung-Stafshede, Pernilla
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