It appears you don't have support to open PDFs in this web browser. To view this file, Open with your PDF reader
Abstract
Protein ubiquitylation is involved in a plethora of cellular processes. While antibodies directed at ubiquitin remnants (K-ɛ-GG) have improved the ability to monitor ubiquitylation using mass spectrometry, methods for highly multiplexed measurement of ubiquitylation in tissues and primary cells using sub-milligram amounts of sample remains a challenge. Here, we present a highly sensitive, rapid and multiplexed protocol termed UbiFast for quantifying ~10,000 ubiquitylation sites from as little as 500 μg peptide per sample from cells or tissue in a TMT10plex in ca. 5 h. High-field Asymmetric Waveform Ion Mobility Spectrometry (FAIMS) is used to improve quantitative accuracy for posttranslational modification analysis. We use the approach to rediscover substrates of the E3 ligase targeting drug lenalidomide and to identify proteins modulated by ubiquitylation in models of basal and luminal human breast cancer. The sensitivity and speed of the UbiFast method makes it suitable for large-scale studies in primary tissue samples.
Comprehensive protein ubiquitylation profiling by mass spectrometry typically requires large sample amounts, limiting its applicability to tissue samples. Here, the authors present an optimized proteomics method that enables multiplexed ubiquitylome analysis of cells and tumor tissue samples.
You have requested "on-the-fly" machine translation of selected content from our databases. This functionality is provided solely for your convenience and is in no way intended to replace human translation. Show full disclaimer
Neither ProQuest nor its licensors make any representations or warranties with respect to the translations. The translations are automatically generated "AS IS" and "AS AVAILABLE" and are not retained in our systems. PROQUEST AND ITS LICENSORS SPECIFICALLY DISCLAIM ANY AND ALL EXPRESS OR IMPLIED WARRANTIES, INCLUDING WITHOUT LIMITATION, ANY WARRANTIES FOR AVAILABILITY, ACCURACY, TIMELINESS, COMPLETENESS, NON-INFRINGMENT, MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE. Your use of the translations is subject to all use restrictions contained in your Electronic Products License Agreement and by using the translation functionality you agree to forgo any and all claims against ProQuest or its licensors for your use of the translation functionality and any output derived there from. Hide full disclaimer
Details





1 Broad Institute of MIT and Harvard, Cambridge, USA (GRID:grid.66859.34)
2 Broad Institute of MIT and Harvard, Cambridge, USA (GRID:grid.66859.34); Brigham and Women’s Hospital, Division of Hematology, Boston, USA (GRID:grid.62560.37) (ISNI:0000 0004 0378 8294); Dana-Farber Cancer Institute, Department of Medical Oncology, Boston, USA (GRID:grid.65499.37) (ISNI:0000 0001 2106 9910)
3 Broad Institute of MIT and Harvard, Cambridge, USA (GRID:grid.66859.34); Brigham and Women’s Hospital, Division of Hematology, Boston, USA (GRID:grid.62560.37) (ISNI:0000 0004 0378 8294); Dana-Farber Cancer Institute, Department of Medical Oncology, Boston, USA (GRID:grid.65499.37) (ISNI:0000 0001 2106 9910); Dana-Farber Cancer Institute, Howard Hughes Medical Institute, Boston, USA (GRID:grid.65499.37) (ISNI:0000 0001 2106 9910)
4 Broad Institute of MIT and Harvard, Cambridge, USA (GRID:grid.66859.34); Max Delbrück Center for Molecular Medicine in the Helmholtz Society, Berlin, Germany (GRID:grid.419491.0) (ISNI:0000 0001 1014 0849); Berlin Institute of Health, Berlin, Germany (GRID:grid.484013.a)