Regulation of Respiration and Apoptosis by Cytochrome c Threonine 58 Phosphorylation

Junmei Wan, Hasini A. Kalpage, Asmita Vaishnav, Jenney Liu, Icksoo Lee, Gargi Mahapatra, Alice A. Turner, Matthew P. Zurek, Qinqin Ji, Carlos T. Moraes, Maurice Andre Recanati, Lawrence I. Grossman, Arthur R. Salomon, Brian F.P. Edwards, Maik Hüttemann

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Cytochrome c (Cytc) is a multifunctional protein, acting as an electron carrier in the electron transport chain (ETC), where it shuttles electrons from bc1 complex to cytochrome c oxidase (COX), and as a trigger of type II apoptosis when released from the mitochondria. We previously showed that Cytc is regulated in a highly tissue-specific manner: Cytc isolated from heart, liver, and kidney is phosphorylated on Y97, Y48, and T28, respectively. Here, we have analyzed the effect of a new Cytc phosphorylation site, threonine 58, which we mapped in rat kidney Cytc by mass spectrometry. We generated and overexpressed wild-type, phosphomimetic T58E, and two controls, T58A and T58I Cytc; the latter replacement is found in human and testis-specific Cytc. In vitro, COX activity, caspase-3 activity, and heme degradation in the presence of H2O2 were decreased with phosphomimetic Cytc compared to wild-type. Cytc-knockout cells expressing T58E or T58I Cytc showed a reduction in intact cell respiration, mitochondrial membrane potential (∆Ψm), ROS production, and apoptotic activity compared to wild-type. We propose that, under physiological conditions, Cytc is phosphorylated, which controls mitochondrial respiration and apoptosis. Under conditions of stress Cytc phosphorylations are lost leading to maximal respiration rates, ∆Ψm hyperpolarization, ROS production, and apoptosis.

Original languageEnglish (US)
Article number15815
JournalScientific reports
Volume9
Issue number1
DOIs
StatePublished - Dec 1 2019
Externally publishedYes

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Threonine
Cytochromes c
Respiration
Phosphorylation
Apoptosis
Electrons
Cell Respiration
Kidney
Mitochondrial Membrane Potential
Electron Transport Complex IV
Respiratory Rate
Electron Transport
Heme
Caspase 3
Testis
Mass Spectrometry
Mitochondria
Oxidoreductases

ASJC Scopus subject areas

  • General

Cite this

Wan, J., Kalpage, H. A., Vaishnav, A., Liu, J., Lee, I., Mahapatra, G., ... Hüttemann, M. (2019). Regulation of Respiration and Apoptosis by Cytochrome c Threonine 58 Phosphorylation. Scientific reports, 9(1), [15815]. https://doi.org/10.1038/s41598-019-52101-z

Regulation of Respiration and Apoptosis by Cytochrome c Threonine 58 Phosphorylation. / Wan, Junmei; Kalpage, Hasini A.; Vaishnav, Asmita; Liu, Jenney; Lee, Icksoo; Mahapatra, Gargi; Turner, Alice A.; Zurek, Matthew P.; Ji, Qinqin; Moraes, Carlos T.; Recanati, Maurice Andre; Grossman, Lawrence I.; Salomon, Arthur R.; Edwards, Brian F.P.; Hüttemann, Maik.

In: Scientific reports, Vol. 9, No. 1, 15815, 01.12.2019.

Research output: Contribution to journalArticle

Wan, J, Kalpage, HA, Vaishnav, A, Liu, J, Lee, I, Mahapatra, G, Turner, AA, Zurek, MP, Ji, Q, Moraes, CT, Recanati, MA, Grossman, LI, Salomon, AR, Edwards, BFP & Hüttemann, M 2019, 'Regulation of Respiration and Apoptosis by Cytochrome c Threonine 58 Phosphorylation', Scientific reports, vol. 9, no. 1, 15815. https://doi.org/10.1038/s41598-019-52101-z
Wan, Junmei ; Kalpage, Hasini A. ; Vaishnav, Asmita ; Liu, Jenney ; Lee, Icksoo ; Mahapatra, Gargi ; Turner, Alice A. ; Zurek, Matthew P. ; Ji, Qinqin ; Moraes, Carlos T. ; Recanati, Maurice Andre ; Grossman, Lawrence I. ; Salomon, Arthur R. ; Edwards, Brian F.P. ; Hüttemann, Maik. / Regulation of Respiration and Apoptosis by Cytochrome c Threonine 58 Phosphorylation. In: Scientific reports. 2019 ; Vol. 9, No. 1.
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AU - Vaishnav, Asmita

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AU - Lee, Icksoo

AU - Mahapatra, Gargi

AU - Turner, Alice A.

AU - Zurek, Matthew P.

AU - Ji, Qinqin

AU - Moraes, Carlos T.

AU - Recanati, Maurice Andre

AU - Grossman, Lawrence I.

AU - Salomon, Arthur R.

AU - Edwards, Brian F.P.

AU - Hüttemann, Maik

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AB - Cytochrome c (Cytc) is a multifunctional protein, acting as an electron carrier in the electron transport chain (ETC), where it shuttles electrons from bc1 complex to cytochrome c oxidase (COX), and as a trigger of type II apoptosis when released from the mitochondria. We previously showed that Cytc is regulated in a highly tissue-specific manner: Cytc isolated from heart, liver, and kidney is phosphorylated on Y97, Y48, and T28, respectively. Here, we have analyzed the effect of a new Cytc phosphorylation site, threonine 58, which we mapped in rat kidney Cytc by mass spectrometry. We generated and overexpressed wild-type, phosphomimetic T58E, and two controls, T58A and T58I Cytc; the latter replacement is found in human and testis-specific Cytc. In vitro, COX activity, caspase-3 activity, and heme degradation in the presence of H2O2 were decreased with phosphomimetic Cytc compared to wild-type. Cytc-knockout cells expressing T58E or T58I Cytc showed a reduction in intact cell respiration, mitochondrial membrane potential (∆Ψm), ROS production, and apoptotic activity compared to wild-type. We propose that, under physiological conditions, Cytc is phosphorylated, which controls mitochondrial respiration and apoptosis. Under conditions of stress Cytc phosphorylations are lost leading to maximal respiration rates, ∆Ψm hyperpolarization, ROS production, and apoptosis.

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