Efficient monitoring of protein ubiquitylation levels using TUBEs-based microarrays

FEBS Lett. 2016 Aug;590(16):2748-56. doi: 10.1002/1873-3468.12289. Epub 2016 Jul 30.


Analyzing protein ubiquitylation changes during physiological or pathological processes is challenging due to its high reversibility and dynamic turnover of modified targets. We have developed a protein microarray to assess endogenous ubiquitylation levels from cell cultures, employing tandem ubiquitin-binding entities (TUBEs) with three or four ubiquitin-associated (UBA) domains as capture probes. Adriamycin (ADR)-stimulated MCF7 cells were used to differentiate protein ubiquitylation levels between cells that are sensitive or resistant to ADR treatment. We show that TUBEs-based microarrays can be used for the analysis of cellular processes regulated by ubiquitylation and for the detection of pathologies with aberrant ubiquitylation levels.

Keywords: Protein microarrays; SPR; TUBEs; UBA domains; ubiquitylation.

Publication types

  • Letter
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Doxorubicin / administration & dosage
  • Humans
  • MCF-7 Cells
  • Protein Array Analysis / methods*
  • Protein Binding / drug effects
  • Proteins / genetics
  • Proteins / metabolism*
  • Ubiquitin / metabolism
  • Ubiquitination / drug effects
  • Ubiquitination / genetics*


  • Proteins
  • Ubiquitin
  • Doxorubicin