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; E. A. De Filippis; R. Zapata Bustos; P. R. Langlais; C. Mielke; L. J. Mandarino

doi:10.1016/j.abb.2018.04.006

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, E. A. De Filippis, R. Zapata Bustos, P. R. Langlais, C. Mielke, L. J. Mandarino

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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 (J_o) 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 J_o 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 language	English (US)
Pages (from-to)	93-103
Number of pages	11
Journal	Archives of Biochemistry and Biophysics
Volume	647
DOIs	https://doi.org/10.1016/j.abb.2018.04.006
State	Published - Jun 1 2018

Keywords

Acetylation
Bioenergetics
Energy metabolism
Flux control

ASJC Scopus subject areas

Biophysics
Biochemistry
Molecular Biology

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10.1016/j.abb.2018.04.006

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Willis, W. T., Miranda-Grandjean, D., Hudgens, J., Willis, E. A., Finlayson, J., De Filippis, E. A., Zapata Bustos, R., Langlais, P. R., Mielke, C., & Mandarino, L. J. (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, 647, 93-103. https://doi.org/10.1016/j.abb.2018.04.006

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

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

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.",

keywords = "Acetylation, Bioenergetics, Energy metabolism, Flux control",

author = "Willis, {W. T.} and D. Miranda-Grandjean and J. Hudgens and Willis, {E. A.} and J. Finlayson and {De Filippis}, {E. A.} and {Zapata Bustos}, R. and Langlais, {P. R.} and C. Mielke and Mandarino, {L. J.}",

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doi = "10.1016/j.abb.2018.04.006",

language = "English (US)",

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T1 - Dominant and sensitive control of oxidative flux by the ATP-ADP carrier in human skeletal muscle mitochondria

T2 - Effect of lysine acetylation

AU - Willis, W. T.

AU - Miranda-Grandjean, D.

AU - Hudgens, J.

AU - Willis, E. A.

AU - Finlayson, J.

AU - De Filippis, E. A.

AU - Zapata Bustos, R.

AU - Langlais, P. R.

AU - Mielke, C.

AU - Mandarino, L. J.

PY - 2018/6/1

Y1 - 2018/6/1

N2 - 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.

AB - 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.

KW - Acetylation

KW - Bioenergetics

KW - Energy metabolism

KW - Flux control

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SN - 0003-9861

VL - 647

SP - 93

EP - 103

JO - Archives of Biochemistry and Biophysics

JF - Archives of Biochemistry and Biophysics

ER -

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

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