Matches in SemOpenAlex for { <https://semopenalex.org/work/W1440773910> ?p ?o ?g. }
- W1440773910 abstract "The ubiquitin pathway is involved in a wide range of cellular events including receptor intemalization, signal transduction and prote as omal degradation. Misregulation of the later can cause many diseases such as cancer and inflammatory disorders. So far, few large-scale studies of protein ubiquitylation have been reported due to the difficulty of isolating these low abundance and rapidly degraded proteins. In this study, a differential proteomic strategy combining affinity purification and the use of a deubiquitylating enzyme (UBP) is proposed to identify ubiquitylated proteins targeted for proteasomal degradation. Two affinity purifications methods were developed, onebased on a protein binding ubiquitylated proteins (Rad23) and the other based on a modified version of ubiquitin (6xHis-ubiquitin). Combination of both purification methods was also evaluated. These purifications methods enabled the identification of more than 1400 potentially ubiquitylated proteins and 28 sites of ubiquitylation with 6 sites on ubiquitin. A differential proteomic approach was then applied by treating half of the enriched ubiquitylated proteins with UBP. The non-treated control and the UBP-treated prote in extracts were digested with trypsin and analyzed by nanoLC/LC-MS/MS on a high resolution LTQ-Orbitrap mass spectrometer. Peptide ions derived from ubiquitylated proteins comprise a miscleavage tryptic site harbouring a characteristic diglycine adduct on the lysine side chain. Comprehensive data mining and clustering analyses on peptide maps using in-house bioinformatics software were performed to characterize two distinct ion populations representing peptides containing or not the diglycine adduct. Validation of the approach enabled the identification of 3 peptide ion pairs correlated with the addition of an ubiquitylation site." @default.
- W1440773910 created "2016-06-24" @default.
- W1440773910 creator A5008437704 @default.
- W1440773910 date "2009-03-05" @default.
- W1440773910 modified "2023-09-23" @default.
- W1440773910 title "Enrichissement de protéines ubiquitinées et une nouvelle approche protéomique pour l'identification des sites d'ubiquitination" @default.
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