, Volume 23, Issue 4, pp 377–388 | Cite as

Protein acetylation on 2-isopropylmalate synthase from Thermus thermophilus HB27

  • Ayako Yoshida
  • Minoru Yoshida
  • Tomohisa Kuzuyama
  • Makoto Nishiyama
  • Saori KosonoEmail author
Original Paper


Protein lysine Nε-acetylation is one of the important factors regulating cellular metabolism. We performed a proteomic analysis to identify acetylated proteins in the extremely thermophilic bacterium, Thermus thermophilus HB27. A total of 335 unique acetylated lysine residues, including many metabolic enzymes and ribosomal proteins, were identified in 208 proteins. Enzymes involved in amino acid metabolism were the most abundant among acetylated metabolic proteins. 2-Isopropylmalate synthase (IPMS), which catalyzes the first step in leucine biosynthesis, was acetylated at four lysine residues. Acetylation-mimicking mutations at Lys332 markedly decreased IPMS activity in vitro, suggesting that Lys332, which is located in subdomain II, plays a regulatory role in IPMS activity. We also investigated the acetylation-deacetylation mechanism of IPMS and revealed that it was acetylated non-enzymatically by acetyl-CoA and deacetylated enzymatically by TT_C0104. The present results suggest that leucine biosynthesis is regulated by post-translational protein modifications, in addition to feedback inhibition/repression, and that metabolic enzymes are regulated by protein acetylation in T. thermophilus.


Proteomics Thermophiles: physiology, metabolism, molecular biology, genetics Protein lysine acetylation 2-Isopropylmalate synthase Leucine biosynthesis Thermus thermophilus 



We thank Kaori Otsuki, Masaya Usui, and Aya Abe at the Support Unit for Bio-Material Analysis, RIKEN-CBS Research Resources Division for their technical assistance with mass spectrometry analyses. This work was supported by JSPS KAKENHI Grant No: JP26870113 and JP17J40083 (AY).

Supplementary material

792_2019_1090_MOESM1_ESM.docx (682 kb)
Supplementary material 1 (DOCX 681 kb)
792_2019_1090_MOESM2_ESM.xlsx (84 kb)
Supplementary material 2 (XLSX 83 kb)


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Copyright information

© Springer Japan KK, part of Springer Nature 2019

Authors and Affiliations

  • Ayako Yoshida
    • 1
  • Minoru Yoshida
    • 2
    • 3
    • 4
  • Tomohisa Kuzuyama
    • 1
    • 4
  • Makoto Nishiyama
    • 1
    • 4
  • Saori Kosono
    • 1
    • 2
    • 4
    Email author
  1. 1.Biotechnology Research CenterThe University of TokyoTokyoJapan
  2. 2.RIKEN Center for Sustainable Resource ScienceWako-shiJapan
  3. 3.Department of Biotechnology, Graduate School of Agricultural and Life SciencesThe University of TokyoTokyoJapan
  4. 4.Collaborative Research Institute for Innovative MicrobiologyThe University of TokyoTokyoJapan

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