Site-specific acetylation of adenine nucleotide translocase 1 at lysine 23 in human muscle

Jean Finlayson, Neusha Barakati, Paul R. Langlais, Janet Funk, Rocio Zapata Bustos, Dawn K. Coletta, Moulun Luo, Wayne T. Willis, Lawrence J. Mandarino

Research output: Contribution to journalArticlepeer-review

1 Scopus citations

Abstract

Evidence suggests acetylation of human adenine nucleotide translocase 1 (ANT1) at lysine 23 (Lys23) reduces binding of ADP. Lys23 contributes to the positive charge that facilitates this interaction. This study was undertaken to characterize ANT1 abundance and acetylation by a novel method using small amounts of human skeletal muscle biopsies. Lysates of whole muscle or mitochondria from the same tissue were prepared from needle biopsies of vastus lateralis muscle of healthy volunteers. Lysed proteins were resolved on gels, the section containing ANT1 (surrounding 30 Kd) was excised, digested with trypsin, spiked with labeled unacetylated and acetylated synthetic standard peptides and analyzed by mass spectrometry. Natural logarithm transformation of data linearized ion intensities over a 10-fold range of peptide mass. Coefficients of variation ranged from 7 to 30% for ANT1 abundance and Lys23 acetylation. In three volunteers, ANT1 content was 8.36 ± 0.33 nmol/g wet weight muscle and 0.64 ± 0.05 nmol/mg mitochondria, so mitochondrial content was 13.3 ± 2.4 mg mitochondria per gram muscle. Acetylation of Lys23 averaged 14.3 ± 4.2% and 4.87 ± 1.84% in whole muscle and mitochondria, respectively. This assay makes it possible to assess effects of acetylation on the function of ANT1 in human muscle.

Original languageEnglish (US)
Article number114319
JournalAnalytical Biochemistry
Volume630
DOIs
StatePublished - Oct 1 2021

Keywords

  • Adenine nucleotide translocase
  • Lysine acetylation
  • Mitochondria
  • Proteomics
  • Skeletal muscle

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Fingerprint

Dive into the research topics of 'Site-specific acetylation of adenine nucleotide translocase 1 at lysine 23 in human muscle'. Together they form a unique fingerprint.

Cite this