Efficient monitoring of protein ubiquitylation levels using TUBEs-based microarrays

Sonia Serna (1) , Wendy Xolalpa (2) , Valérie Lang (2) , Fabienne Aillet (2) , Patrick England (3) , Niels Reichardt (1, 4) , Manuel S Rodriguez (5, 2, 6)

1 CIC biomaGUNE - Centro de Investigación Cooperativa en Biomateriales
2 Inbiomed [Spain]
3 IP - Institut Pasteur [Paris]
4 CIBER-BBN - Centro de Investigación Biomédica en Red en Bioingeniería, Biomateriales y Nanomedicina
5 ITAV - Institut des Technologies Avancées en sciences du Vivant
6 IPBS - Institut de pharmacologie et de biologie structurale

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

Domains: Life Sciences [q-bio]

Sonia Serna, Wendy Xolalpa, Valérie Lang, Fabienne Aillet, Patrick England, et al.. Efficient monitoring of protein ubiquitylation levels using TUBEs-based microarrays. FEBS Letters, 2016, 590 (16), pp.2748 - 2756. ⟨10.1002/1873-3468.12289⟩. ⟨pasteur-03525440⟩