title: Efficient monitoring of protein ubiquitylation levels using TUBEs-based microarrays.
abstract: 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.
authors.affiliation: Glycotechnology Laboratory, CIC biomaGUNE, San Sebastián-Donostia, Spain.
authors.forename: Sonia
authors.initials: S
authors.lastname: Serna
authors.affiliation: Ubiquitylation and Cancer Molecular Biology Laboratory, Inbiomed, San Sebastián-Donostia, Spain.
authors.forename: Wendy
authors.initials: W
authors.lastname: Xolalpa
authors.affiliation: Ubiquitylation and Cancer Molecular Biology Laboratory, Inbiomed, San Sebastián-Donostia, Spain.
authors.forename: Valérie
authors.initials: V
authors.lastname: Lang
authors.affiliation: Ubiquitylation and Cancer Molecular Biology Laboratory, Inbiomed, San Sebastián-Donostia, Spain.
authors.forename: Fabienne
authors.initials: F
authors.lastname: Aillet
authors.affiliation: Institut Pasteur, Molecular Biophysics Facility, Citech and Department of Structural Biology and Chemistry, Paris, France.|CNRS UMR3528, Paris, France.
authors.forename: Patrick
authors.initials: P
authors.lastname: England
authors.affiliation: Glycotechnology Laboratory, CIC biomaGUNE, San Sebastián-Donostia, Spain.|CIBER de Bioingeniería, Biomateriales y Nanomedicina (CIBER-BBN), San Sebastian, Spain.
authors.forename: Niels
authors.initials: N
authors.lastname: Reichardt
authors.affiliation: Ubiquitylation and Cancer Molecular Biology Laboratory, Inbiomed, San Sebastián-Donostia, Spain.|Institut des Technologies Avancées en sciences du Vivant (ITAV), Université de Toulouse, CNRS, UPS, France.|Institut de Pharmacologie et de Biologie Structurale (IPBS), Université de Toulouse, CNRS, UPS, France.
authors.forename: Manuel S
authors.initials: MS
authors.lastname: Rodriguez
mesh_terms: D004317:Doxorubicin / Q000008:administration & dosage; D006801:Humans; D061986:MCF-7 Cells; D040081:Protein Array Analysis / Q000379:methods*; D011485:Protein Binding / Q000187:drug effects; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D025801:Ubiquitin / Q000378:metabolism; D054875:Ubiquitination / Q000187:drug effects / Q000235:genetics*
chemical_list: D011506:Proteins; D025801:Ubiquitin; D004317:Doxorubicin
keywords: Protein microarrays; SPR; TUBEs; UBA domains; ubiquitylation