Dominant and sensitive control of oxidative flux by the ATP-ADP carrier in human skeletal muscle mitochondria: Effect of lysine acetylation

W. T. Willis, D. Miranda-Grandjean, J. Hudgens, E. A. Willis, J. Finlayson, Elena Anna De Filippis, R. Zapata Bustos, Paul R Langlais, C. Mielke, L. J. Mandarino

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

The adenine nucleotide translocase (ANT) of the mitochondrial inner membrane exchanges ADP for ATP. Mitochondria were isolated from human vastus lateralis muscle (n = 9). Carboxyatractyloside titration of O2 consumption rate (Jo) at clamped [ADP] of 21 μM gave ANT abundance of 0.97 ± 0.14 nmol ANT/mg and a flux control coefficient of 82% ± 6%. Flux control fell to 1% ± 1% at saturating (2 mM) [ADP]. The KmADP for Jo was 32.4 ± 1.8 μM. In terms of the free (−3) ADP anion this KmADP was 12.0 ± 0.7 μM. A novel luciferase-based assay for ATP production gave KmADP of 13.1 ± 1.9 μM in the absence of ATP competition. The free anion KmADP in this case was 2.0 ± 0.3 μM. Targeted proteomic analyses showed significant acetylation of ANT Lysine23 and that ANT1 was the most abundant isoform. Acetylation of Lysine23 correlated positively with KmADP, r = 0.74, P = 0.022. The findings underscore the central role played by ANT in the control of oxidative phosphorylation, particularly at the energy phosphate levels associated with low ATP demand. As predicted by molecular dynamic modeling, ANT Lysine23 acetylation decreased the apparent affinity of ADP for ANT binding.

Original languageEnglish (US)
Pages (from-to)93-103
Number of pages11
JournalArchives of Biochemistry and Biophysics
Volume647
DOIs
StatePublished - Jun 1 2018

Fingerprint

ATP Translocases Mitochondrial ADP
Muscle Mitochondrion
Acetylation
Mitochondria
Lysine
Muscle
Skeletal Muscle
Fluxes
Adenosine Diphosphate
Adenosine Triphosphate
Anions
Oxidative Phosphorylation
Quadriceps Muscle
Mitochondrial Membranes
Molecular Dynamics Simulation
Luciferases
Titration
Proteomics
Molecular dynamics
Assays

Keywords

  • Acetylation
  • Bioenergetics
  • Energy metabolism
  • Flux control

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology

Cite this

Dominant and sensitive control of oxidative flux by the ATP-ADP carrier in human skeletal muscle mitochondria : Effect of lysine acetylation. / Willis, W. T.; Miranda-Grandjean, D.; Hudgens, J.; Willis, E. A.; Finlayson, J.; De Filippis, Elena Anna; Zapata Bustos, R.; Langlais, Paul R; Mielke, C.; Mandarino, L. J.

In: Archives of Biochemistry and Biophysics, Vol. 647, 01.06.2018, p. 93-103.

Research output: Contribution to journalArticle

Willis, WT, Miranda-Grandjean, D, Hudgens, J, Willis, EA, Finlayson, J, De Filippis, EA, Zapata Bustos, R, Langlais, PR, Mielke, C & Mandarino, LJ 2018, 'Dominant and sensitive control of oxidative flux by the ATP-ADP carrier in human skeletal muscle mitochondria: Effect of lysine acetylation', Archives of Biochemistry and Biophysics, vol. 647, pp. 93-103. https://doi.org/10.1016/j.abb.2018.04.006
Willis, W. T. ; Miranda-Grandjean, D. ; Hudgens, J. ; Willis, E. A. ; Finlayson, J. ; De Filippis, Elena Anna ; Zapata Bustos, R. ; Langlais, Paul R ; Mielke, C. ; Mandarino, L. J. / Dominant and sensitive control of oxidative flux by the ATP-ADP carrier in human skeletal muscle mitochondria : Effect of lysine acetylation. In: Archives of Biochemistry and Biophysics. 2018 ; Vol. 647. pp. 93-103.
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AU - Willis, E. A.

AU - Finlayson, J.

AU - De Filippis, Elena Anna

AU - Zapata Bustos, R.

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