| |  Online-Ressource |
|---|
| Verfasst von: | Zinn, Nico [VerfasserIn]  |
|---|
| | Werner, Thilo [VerfasserIn] |
|---|
| | Doce, Carola [VerfasserIn] |
|---|
| | Mathieson, Toby [VerfasserIn] |
|---|
| | Böcker, Christine [VerfasserIn] |
|---|
| | Sweetman, Gavain [VerfasserIn] |
|---|
| | Fufezan, Christian [VerfasserIn] |
|---|
| | Bantscheff, Marcus [VerfasserIn] |
|---|
| Titel: | Improved proteomics-based drug mechanism-of-action studies using 16-plex isobaric mass tags |
|---|
| Verf.angabe: | Nico Zinn, Thilo Werner, Carola Doce, Toby Mathieson, Christine Boecker, Gavain Sweetman, Christian Fufezan, and Marcus Bantscheff |
|---|
| E-Jahr: | 2021 |
|---|
| Jahr: | February 23, 2021 |
|---|
| Umfang: | 10 S. |
|---|
| Teil: | volume:20 |
|---|
| | year:2021 |
|---|
| | number:3 |
|---|
| | pages:1792-1801 |
|---|
| | extent:10 |
|---|
| Fussnoten: | Gesehen am 04.05.2021 |
|---|
| Titel Quelle: | Enthalten in: Journal of proteome research |
|---|
| Ort Quelle: | Washington, DC : ACS Publications, 2002 |
|---|
| Jahr Quelle: | 2021 |
|---|
| Band/Heft Quelle: | 20(2021), 3, Seite 1792-1801 |
|---|
| ISSN Quelle: | 1535-3907 |
|---|
| Abstract: | Multiplexed quantitative proteomics enabled complex workflows to study the mechanisms by which small molecule drugs interact with the proteome such as thermal proteome profiling (TPP) or multiplexed proteome dynamics profiling (mPDP). TPP measures changes in protein thermal stability in response to drug treatment and thus informs on direct targets and downstream regulation events, while the mPDP approach enables the discovery of regulated protein synthesis and degradation events caused by small molecules and other perturbations. The isobaric mass tags available for multiplexed proteomics have thus far limited the efficiency and sensitivity by which such experiments could be performed. Here we evaluate a recent generation of 16-plex isobaric mass tags and demonstrate the sensitive and time efficient identification of Staurosporine targets in HepG2 cell extracts by recording full thermal denaturation/aggregation profiles of vehicle and compound treated samples in a single mass spectrometry experiment. In 2D-TPP experiments, isothermal titration over seven concentrations per temperature enabled comprehensive selectivity profiling of Staurosporine with EC50 values for kinase targets tightly matching to the kinobeads gold standard assay. Finally, we demonstrate time and condition-based multiplexing of dynamic SILAC labeling experiments to delineate proteome-wide effects of the molecular glue Indisulam on synthesis and degradation rates. |
|---|
| DOI: | doi:10.1021/acs.jproteome.0c00900 |
|---|
| URL: | Bitte beachten Sie: Dies ist ein Bibliographieeintrag. Ein Volltextzugriff für Mitglieder der Universität besteht hier nur, falls für die entsprechende Zeitschrift/den entsprechenden Sammelband ein Abonnement besteht oder es sich um einen OpenAccess-Titel handelt.
Volltext: https://doi.org/10.1021/acs.jproteome.0c00900 |
|---|
| | DOI: https://doi.org/10.1021/acs.jproteome.0c00900 |
|---|
| Datenträger: | Online-Ressource |
|---|
| Sprache: | eng |
|---|
| K10plus-PPN: | 1757077367 |
|---|
| Verknüpfungen: | → Zeitschrift |
|---|
Improved proteomics-based drug mechanism-of-action studies using 16-plex isobaric mass tags / Zinn, Nico [VerfasserIn]; February 23, 2021 (Online-Ressource)
