The putative leucine sensor Sestrin2 is hyperphosphorylated by acute resistance exercise but not protein ingestion in human skeletal muscle

  • Nina Zeng
  • Randall F. D’Souza
  • Brie Sorrenson
  • Troy L. Merry
  • Matthew P. G. Barnett
  • Cameron J. Mitchell
  • David Cameron-Smith
Original Article

Abstract

Purpose

Dietary protein and resistance exercise (RE) are both potent stimuli of the mammalian target of rapamycin complex 1 (mTORC1). Sestrins1, 2, 3 are multifunctional proteins that regulate mTORC1, stimulate autophagy and alleviate oxidative stress. Of this family, Sestrin2 is a putative leucine sensor implicated in mTORC1 and AMP-dependent protein kinase (AMPK) regulation. There is currently no data examining the responsiveness of Sestrin2 to dietary protein ingestion, with or without RE.

Methods

In Study 1, 16 males ingested either 10 or 20 g of milk protein concentrate (MPC) with muscle biopsies collected pre, 90 and 210 min post-beverage consumption. In Study 2, 20 males performed a bout of RE immediately followed by the consumption of 9 g of MPC or carbohydrate placebo. Analysis of Sestrins, AMPK and antioxidant responses was examined.

Results

Dietary protein ingestion did not result in Sestrin2 mobility shift. After RE, Sestrin2 phosphorylation state was significantly altered and was not further modified by post-exercise protein or carbohydrate ingestion. With RE, AMPK phosphorylation remained stable, while the mRNA expressions of several antioxidants were upregulated.

Conclusions

Dietary protein ingestion did not affect the signalling by the family of Sestrins. With RE, Sestrin2 was hyperphosphorylated, with no further evidence of a relationship to AMPK signalling.

Keywords

Amino acid Resistance exercise Mammalian target of rapamycin Oxidative stress Antioxidant Sestrins Hyperphosphorylation 

Abbreviations

4E BP1

Eukaryotic initiation factor 4E-binding protein 1

AA

Amino acid

AMPK

AMP-activated protein kinase

ANOVA

Analysis of variance

CHO

Carbohydrate

GATOR

GTPase-activating protein activity towards Rags

GCLM

Glutamate-cysteine ligase

HEK293

Human embryonic kidney cells 293

HMOX1

Heme oxygenase 1

Keap1

Kelch-like ECH-associated protein

MPC

Milk protein concentrate

MPS

Muscle protein synthesis

mRNA

Messenger RNA

mTORC1

Mammalian target of rapamycin complex 1

NFE2L2

Nuclear factor (erythroid-derived 2)-like 2

p70S6K1

Ribosomal protein S6 kinase 1

p62

Sequestosome 1

RE

Resistance exercise

RM

Repetition maximum

ROS

Reactive oxygen species

RPS6

Ribosomal protein S6

SEM

Standard error of mean

SESN

Sestrin

SRXN1

Sulfiredoxin-1

Notes

Acknowledgements

The study was funded by AgResearch (through Strategic Science Investment Fund contracts A19079 and A21246). MPGB is a current employee of AgResearch.

Author contributions

NZ, CJM and DCS designed the study; NZ and RFD conducted experiments; NZ, BS and TLM analysed data; NZ, CJM and DCS drafted manuscript; NZ, RFD, MPGB, BS, TLM, CJM and DCS critically evaluated and contributed to the manuscript.

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Nina Zeng
    • 1
  • Randall F. D’Souza
    • 1
  • Brie Sorrenson
    • 2
    • 3
  • Troy L. Merry
    • 3
    • 4
  • Matthew P. G. Barnett
    • 5
  • Cameron J. Mitchell
    • 1
  • David Cameron-Smith
    • 1
    • 6
    • 7
  1. 1.Liggins InstituteThe University of AucklandAucklandNew Zealand
  2. 2.Department of Molecular Medicine and Pathology, Faculty of Medical and Health SciencesThe University of AucklandAucklandNew Zealand
  3. 3.Maurice Wilkins Centre for Molecular BiodiscoveryThe University of AucklandAucklandNew Zealand
  4. 4.Discipline of Nutrition, Faculty of Medical and Health SciencesThe University of AucklandAucklandNew Zealand
  5. 5.Food Nutrition and Health TeamAgResearchPalmerston NorthNew Zealand
  6. 6.Food and Bio-based Products GroupAgResearchPalmerston NorthNew Zealand
  7. 7.Riddet InstitutePalmerston NorthNew Zealand

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