The activity of Menkes disease protein ATP7A is essential for redox balance in mitochondria

Ashima Bhattacharjee, Haojun Yang, Megan Duffy, Emily Robinson, Arianrhod Conrad-Antoville, Ya Wen Lu, Tony Capps, Lelita Braiterman, Michael Wolfgang, Michael P. Murphy, Ling Yi, Stephen G. Kaler, Svetlana Lutsenko, Martina Ralle

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Copper-transporting ATPase ATP7A is essential for mammalian copper homeostasis. Loss of ATP7A activity is associated with fatal Menkes disease and various other pathologies. In cells, ATP7A inactivation disrupts copper transport from the cytosol into the secretory pathway. Using fibroblasts from Menkes disease patients and mouse 3T3-L1 cells with a CRISPR/Cas9-inactivated ATP7A, we demonstrate that ATP7A dysfunction is also damaging to mitochondrial redox balance. In these cells, copper accumulates in nuclei, cytosol, and mitochondria, causing distinct changes in their redox environment. Quantitative imaging of live cells using GRX1-roGFP2 and HyPer sensors reveals highest glutathione oxidation and elevation of H2O2 in mitochondria, whereas the redox environment of nuclei and the cytosol is much less affected. Decreasing the H2O2 levels in mitochondria with MitoQ does not prevent glutathione oxidation; i.e. elevated copper and not H2O2 is a primary cause of glutathione oxidation. Redox misbalance does not significantly affect mitochondrion morphology or the activity of respiratory complex IV but markedly increases cell sensitivity to even mild glutathione depletion, resulting in loss of cell viability. Thus, ATP7A activity protects mitochondria from excessive copper entry, which is deleterious to redox buffers. Mitochondrial redox misbalance could significantly contribute to pathologies associated with ATP7A inactivation in tissues with paradoxical accumulation of copper (i.e. renal epithelia).

Original languageEnglish (US)
Pages (from-to)16644-16658
Number of pages15
JournalJournal of Biological Chemistry
Volume291
Issue number32
DOIs
StatePublished - Aug 5 2016

Fingerprint

Menkes Kinky Hair Syndrome
Mitochondria
Oxidation-Reduction
Copper
Glutathione
Cytosol
Proteins
Pathology
Oxidation
Clustered Regularly Interspaced Short Palindromic Repeats
3T3-L1 Cells
Secretory Pathway
Fibroblasts
Cell Survival
Buffers
Homeostasis
Epithelium
Cells
Tissue
Kidney

ASJC Scopus subject areas

  • Biochemistry
  • Medicine(all)
  • Molecular Biology
  • Cell Biology

Cite this

Bhattacharjee, A., Yang, H., Duffy, M., Robinson, E., Conrad-Antoville, A., Lu, Y. W., ... Ralle, M. (2016). The activity of Menkes disease protein ATP7A is essential for redox balance in mitochondria. Journal of Biological Chemistry, 291(32), 16644-16658. https://doi.org/10.1074/jbc.M116.727248

The activity of Menkes disease protein ATP7A is essential for redox balance in mitochondria. / Bhattacharjee, Ashima; Yang, Haojun; Duffy, Megan; Robinson, Emily; Conrad-Antoville, Arianrhod; Lu, Ya Wen; Capps, Tony; Braiterman, Lelita; Wolfgang, Michael; Murphy, Michael P.; Yi, Ling; Kaler, Stephen G.; Lutsenko, Svetlana; Ralle, Martina.

In: Journal of Biological Chemistry, Vol. 291, No. 32, 05.08.2016, p. 16644-16658.

Research output: Contribution to journalArticle

Bhattacharjee, A, Yang, H, Duffy, M, Robinson, E, Conrad-Antoville, A, Lu, YW, Capps, T, Braiterman, L, Wolfgang, M, Murphy, MP, Yi, L, Kaler, SG, Lutsenko, S & Ralle, M 2016, 'The activity of Menkes disease protein ATP7A is essential for redox balance in mitochondria', Journal of Biological Chemistry, vol. 291, no. 32, pp. 16644-16658. https://doi.org/10.1074/jbc.M116.727248
Bhattacharjee A, Yang H, Duffy M, Robinson E, Conrad-Antoville A, Lu YW et al. The activity of Menkes disease protein ATP7A is essential for redox balance in mitochondria. Journal of Biological Chemistry. 2016 Aug 5;291(32):16644-16658. https://doi.org/10.1074/jbc.M116.727248
Bhattacharjee, Ashima ; Yang, Haojun ; Duffy, Megan ; Robinson, Emily ; Conrad-Antoville, Arianrhod ; Lu, Ya Wen ; Capps, Tony ; Braiterman, Lelita ; Wolfgang, Michael ; Murphy, Michael P. ; Yi, Ling ; Kaler, Stephen G. ; Lutsenko, Svetlana ; Ralle, Martina. / The activity of Menkes disease protein ATP7A is essential for redox balance in mitochondria. In: Journal of Biological Chemistry. 2016 ; Vol. 291, No. 32. pp. 16644-16658.
@article{f4327f9906cf46049609b644fa0daaaa,
title = "The activity of Menkes disease protein ATP7A is essential for redox balance in mitochondria",
abstract = "Copper-transporting ATPase ATP7A is essential for mammalian copper homeostasis. Loss of ATP7A activity is associated with fatal Menkes disease and various other pathologies. In cells, ATP7A inactivation disrupts copper transport from the cytosol into the secretory pathway. Using fibroblasts from Menkes disease patients and mouse 3T3-L1 cells with a CRISPR/Cas9-inactivated ATP7A, we demonstrate that ATP7A dysfunction is also damaging to mitochondrial redox balance. In these cells, copper accumulates in nuclei, cytosol, and mitochondria, causing distinct changes in their redox environment. Quantitative imaging of live cells using GRX1-roGFP2 and HyPer sensors reveals highest glutathione oxidation and elevation of H2O2 in mitochondria, whereas the redox environment of nuclei and the cytosol is much less affected. Decreasing the H2O2 levels in mitochondria with MitoQ does not prevent glutathione oxidation; i.e. elevated copper and not H2O2 is a primary cause of glutathione oxidation. Redox misbalance does not significantly affect mitochondrion morphology or the activity of respiratory complex IV but markedly increases cell sensitivity to even mild glutathione depletion, resulting in loss of cell viability. Thus, ATP7A activity protects mitochondria from excessive copper entry, which is deleterious to redox buffers. Mitochondrial redox misbalance could significantly contribute to pathologies associated with ATP7A inactivation in tissues with paradoxical accumulation of copper (i.e. renal epithelia).",
author = "Ashima Bhattacharjee and Haojun Yang and Megan Duffy and Emily Robinson and Arianrhod Conrad-Antoville and Lu, {Ya Wen} and Tony Capps and Lelita Braiterman and Michael Wolfgang and Murphy, {Michael P.} and Ling Yi and Kaler, {Stephen G.} and Svetlana Lutsenko and Martina Ralle",
year = "2016",
month = "8",
day = "5",
doi = "10.1074/jbc.M116.727248",
language = "English (US)",
volume = "291",
pages = "16644--16658",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "32",

}

TY - JOUR

T1 - The activity of Menkes disease protein ATP7A is essential for redox balance in mitochondria

AU - Bhattacharjee, Ashima

AU - Yang, Haojun

AU - Duffy, Megan

AU - Robinson, Emily

AU - Conrad-Antoville, Arianrhod

AU - Lu, Ya Wen

AU - Capps, Tony

AU - Braiterman, Lelita

AU - Wolfgang, Michael

AU - Murphy, Michael P.

AU - Yi, Ling

AU - Kaler, Stephen G.

AU - Lutsenko, Svetlana

AU - Ralle, Martina

PY - 2016/8/5

Y1 - 2016/8/5

N2 - Copper-transporting ATPase ATP7A is essential for mammalian copper homeostasis. Loss of ATP7A activity is associated with fatal Menkes disease and various other pathologies. In cells, ATP7A inactivation disrupts copper transport from the cytosol into the secretory pathway. Using fibroblasts from Menkes disease patients and mouse 3T3-L1 cells with a CRISPR/Cas9-inactivated ATP7A, we demonstrate that ATP7A dysfunction is also damaging to mitochondrial redox balance. In these cells, copper accumulates in nuclei, cytosol, and mitochondria, causing distinct changes in their redox environment. Quantitative imaging of live cells using GRX1-roGFP2 and HyPer sensors reveals highest glutathione oxidation and elevation of H2O2 in mitochondria, whereas the redox environment of nuclei and the cytosol is much less affected. Decreasing the H2O2 levels in mitochondria with MitoQ does not prevent glutathione oxidation; i.e. elevated copper and not H2O2 is a primary cause of glutathione oxidation. Redox misbalance does not significantly affect mitochondrion morphology or the activity of respiratory complex IV but markedly increases cell sensitivity to even mild glutathione depletion, resulting in loss of cell viability. Thus, ATP7A activity protects mitochondria from excessive copper entry, which is deleterious to redox buffers. Mitochondrial redox misbalance could significantly contribute to pathologies associated with ATP7A inactivation in tissues with paradoxical accumulation of copper (i.e. renal epithelia).

AB - Copper-transporting ATPase ATP7A is essential for mammalian copper homeostasis. Loss of ATP7A activity is associated with fatal Menkes disease and various other pathologies. In cells, ATP7A inactivation disrupts copper transport from the cytosol into the secretory pathway. Using fibroblasts from Menkes disease patients and mouse 3T3-L1 cells with a CRISPR/Cas9-inactivated ATP7A, we demonstrate that ATP7A dysfunction is also damaging to mitochondrial redox balance. In these cells, copper accumulates in nuclei, cytosol, and mitochondria, causing distinct changes in their redox environment. Quantitative imaging of live cells using GRX1-roGFP2 and HyPer sensors reveals highest glutathione oxidation and elevation of H2O2 in mitochondria, whereas the redox environment of nuclei and the cytosol is much less affected. Decreasing the H2O2 levels in mitochondria with MitoQ does not prevent glutathione oxidation; i.e. elevated copper and not H2O2 is a primary cause of glutathione oxidation. Redox misbalance does not significantly affect mitochondrion morphology or the activity of respiratory complex IV but markedly increases cell sensitivity to even mild glutathione depletion, resulting in loss of cell viability. Thus, ATP7A activity protects mitochondria from excessive copper entry, which is deleterious to redox buffers. Mitochondrial redox misbalance could significantly contribute to pathologies associated with ATP7A inactivation in tissues with paradoxical accumulation of copper (i.e. renal epithelia).

UR - http://www.scopus.com/inward/record.url?scp=84982812274&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84982812274&partnerID=8YFLogxK

U2 - 10.1074/jbc.M116.727248

DO - 10.1074/jbc.M116.727248

M3 - Article

C2 - 27226607

AN - SCOPUS:84982812274

VL - 291

SP - 16644

EP - 16658

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 32

ER -