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Efficient monitoring of protein ubiquitylation levels using TUBE s‐based microarrays
Author(s) -
Serna Sonia,
Xolalpa Wendy,
Lang Valérie,
Aillet Fabienne,
England Patrick,
Reichardt Niels,
Rodriguez Manuel S.
Publication year - 2016
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1002/1873-3468.12289
Subject(s) - ubiquitin , microarray , protein microarray , endogeny , dna microarray , microbiology and biotechnology , chemistry , biology , ubiquitin ligase , biochemistry , gene expression , gene
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 ( TUBE s) with three or four ubiquitin‐associated ( UBA ) domains as capture probes. Adriamycin ( ADR )‐stimulated MCF 7 cells were used to differentiate protein ubiquitylation levels between cells that are sensitive or resistant to ADR treatment. We show that TUBE s‐based microarrays can be used for the analysis of cellular processes regulated by ubiquitylation and for the detection of pathologies with aberrant ubiquitylation levels.