Biochemistry (Moscow)

, Volume 82, Issue 9, pp 1042–1047 | Cite as

Quantitative affinity interaction of ubiquitinated and non-ubiquitinated proteins with proteasome subunit Rpn10

  • O. A. Buneeva
  • O. V. Gnedenko
  • A. T. Kopylov
  • M. V. Medvedeva
  • V. G. Zgoda
  • A. S. Ivanov
  • A. E. MedvedevEmail author


Recent proteomic profiling of mouse brain preparations using the ubiquitin receptor, Rpn10 proteasome subunit, as an affinity ligand revealed a representative group of proteins bound to this sorbent (Medvedev, A. E., et al. (2017) Biochemistry (Moscow), 82, 330-339). In the present study, we investigated interaction of the Rpn10 subunit of proteasomes with some of these identified proteins: glyceraldehyde-3-phosphate dehydrogenase (GAPDH), pyruvate kinase, and histones H2A and H2B. The study revealed: (i) quantitative affinity interaction of the proteasome subunit immobilized on a Biacore-3000 optical biosensor cuvette with both the GAPDH (K d = 2.4·10–6 M) and pyruvate kinase (K d = 2.8·10–5 M); (ii) quantitative high-affinity interaction of immobilized histones H2A and H2B with the Rpn10 subunit (Kd values of 6.5·10–8 and 3.2·10–9 M, respectively). Mass spectrometric analysis revealed the presence of the ubiquitin signature (GG) only in a highly purified preparation of GAPDH. We suggest that binding (especially high-affinity binding) of non-ubiquitinated proteins to the Rpn10 proteasome subunit can both regulate the functioning of this proteasomal ubiquitin receptor (by competing with ubiquitinated substrates) and promote activation of other pathways for proteolytic degradation of proteins destined to the proteasome.


Rpn10 proteasome subunit Rpn10-binding proteins optical biosensor ubiquitin signature 



glyceraldehyde-3-phosphate dehydrogenase


equilibrium dissociation constant


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Copyright information

© Pleiades Publishing, Ltd. 2017

Authors and Affiliations

  • O. A. Buneeva
    • 1
  • O. V. Gnedenko
    • 1
  • A. T. Kopylov
    • 1
  • M. V. Medvedeva
    • 2
  • V. G. Zgoda
    • 1
  • A. S. Ivanov
    • 1
  • A. E. Medvedev
    • 1
    Email author
  1. 1.Orekhovich Institute of Biomedical ChemistryMoscowRussia
  2. 2.Lomonosov Moscow State UniversityBiological Faculty, o119991MoscowRussia

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