The N-terminal shuttle domain of Erv1 determines the affinity for Mia40 and mediates electron transfer to the catalytic Erv1 core in yeast mitochondria

Lionaki, E., Aivaliotis, M., Pozidis, C. and Tokatlidis, K. (2010) The N-terminal shuttle domain of Erv1 determines the affinity for Mia40 and mediates electron transfer to the catalytic Erv1 core in yeast mitochondria. Antioxidants and Redox Signaling, 13(9), pp. 1327-1339. (doi:10.1089/ars.2010.3200)

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Abstract

Erv1 and Mia40 constitute the two important components of the disulfide relay system that mediates oxidative protein folding in the mitochondrial intermembrane space. Mia40 is the import receptor that recognizes the substrates introducing disulfide bonds while it is reduced. A key function of Erv1 is to recycle Mia40 to its active oxidative state. Our aims here were to dissect the domain of Erv1 that mediates the protein–protein interaction with Mia40 and to investigate the interactions between the shuttle domain of Erv1 and its catalytic core and their relevance for the interaction with Mia40. We purified these domains separately as well as cysteine mutants in the shuttle and the active core domains. The noncovalent interaction of Mia40 with Erv1 was measured by isothermal titration calorimetry, whereas their covalent mixed disulfide intermediate was analyzed in reconstitution experiments in vitro and in organello. We established that the N-terminal shuttle domain of Erv1 is necessary and sufficient for interaction to occur. Furthermore, we provide direct evidence for the intramolecular electron transfer from the shuttle cysteine pair of Erv1 to the core domain. Finally, we reconstituted the system by adding in trans the N- and C- terminal domains of Erv1 together with its substrate Mia40.

Item Type:Articles
Additional Information:This is a copy of an article published in the Antioxidants and Redox Signaling © 2010 copyright Mary Ann Liebert, Inc.; Antioxidants and Redox Signaling is available online at: http://online.liebertpub.com.
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Tokatlidis, Professor Kostas
Authors: Lionaki, E., Aivaliotis, M., Pozidis, C., and Tokatlidis, K.
College/School:College of Medical Veterinary and Life Sciences > Institute of Molecular Cell and Systems Biology
Journal Name:Antioxidants and Redox Signaling
Publisher:Mary Ann Liebert, Inc.
ISSN:1523-0864
ISSN (Online):1557-7716
Copyright Holders:Copyright © 2010 Mary Ann Liebert, Inc.
First Published:First published in Antioxidants and Redox Signaling 13(9):1327-1339
Publisher Policy:Reproduced in accordance with the copyright policy of the publisher

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