Mitochondrial copper(I) transfer from Cox17 to Sco1 is coupled to electron transfer

doi:10.1073/pnas.0800019105

Mitochondrial copper(I) transfer from Cox17 to Sco1 is coupled to electron transfer

*Magnetic Resonance Center (CERM) and Department of Chemistry, University of Florence, Via Luigi Sacconi 6, 50019 Sesto Fiorentino, Florence, Italy; and
^‡Department of Gene Technology, Tallinn University of Technology, Akadeemia tee 15, 12618 Tallinn, Estonia

Edited by Harry B. Gray, California Institute of Technology, Pasadena, CA, and approved February 13, 2008 (received for review January 2, 2008)

Abstract

The human protein Cox17 contains three pairs of cysteines. In the mitochondrial intermembrane space (IMS) it exists in a partially oxidized form with two S–S bonds and two reduced cysteines (HCox17_2S-S). HCox17_2S-S is involved in copper transfer to the human cochaperones Sco1 and Cox11, which are implicated in the assembly of cytochrome c oxidase. We show here that Cu(I)HCox17_2S-S, i.e., the copper-loaded form of the protein, can transfer simultaneously copper(I) and two electrons to the human cochaperone Sco1 (HSco1) in the oxidized state, i.e., with its metal-binding cysteines forming a disulfide bond. The result is Cu(I)HSco1 and the fully oxidized apoHCox17_3S-S, which can be then reduced by glutathione to apoHCox17_2S-S. The HSco1/HCox17_2S-S redox reaction is thermodynamically driven by copper transfer. These reactions may occur in vivo because HSco1 can be found in the partially oxidized state within the IMS, consistent with the variable redox properties of the latter compartment. The electron transfer-coupled metallation of HSco1 can be a mechanism within the IMS for an efficient specific transfer of the metal to proteins, where metal-binding thiols are oxidized. The same reaction of copper–electron-coupled transfer does not occur with the human homolog of Sco1, HSco2, for kinetic reasons that may be ascribed to the lack of a specific metal-bridged protein–protein complex, which is instead observed in the Cu(I)HCox17_2S-S/HSco1 interaction.

Footnotes

^†To whom correspondence should be addressed. E-mail: ivanobertini{at}cerm.unifi.it
Author contributions: L.B. and I.B. designed research; S.C.-B., T.H., M.M., and P.P. performed research; S.C.-B., T.H., M.M., and P.P. analyzed data; and L.B., I.B., S.C.-B., and P.P. wrote the paper.
The authors declare no conflict of interest.
This article is a PNAS Direct Submission.
This article contains supporting information online at www.pnas.org/cgi/content/full/0800019105/DCSupplemental.
↵§ When, to a 1:1 mixture of ¹⁵N-labeled apoHSco1 and ¹⁵N-labeled apoHSco2, 1 eq of a copper(I) acetonitrile complex was added, formation of the copper form for the two proteins was observed in similar ratios, thus indicating similar cooper(I)-binding affinity (Fig. S4); the copper forms are in a slow exchange on the chemical shift time scale with the apo forms.