Comparison of Quantitative Mass Spectrometric Methods for Drug Target Identification by Thermal Proteome Profiling.
J Proteome Res
; 22(8): 2629-2640, 2023 08 04.
Article
in En
| MEDLINE
| ID: mdl-37439223
ABSTRACT
Thermal proteome profiling (TPP) provides a powerful approach to studying proteome-wide interactions of small therapeutic molecules and their target and off-target proteins, complementing phenotypic-based drug screens. Detecting differences in thermal stability due to target engagement requires high quantitative accuracy and consistent detection. Isobaric tandem mass tags (TMTs) are used to multiplex samples and increase quantification precision in TPP analysis by data-dependent acquisition (DDA). However, advances in data-independent acquisition (DIA) can provide higher sensitivity and protein coverage with reduced costs and sample preparation steps. Herein, we explored the performance of different DIA-based label-free quantification approaches compared to TMT-DDA for thermal shift quantitation. Acute myeloid leukemia cells were treated with losmapimod, a known inhibitor of MAPK14 (p38α). Label-free DIA approaches, and particularly the library-free mode in DIA-NN, were comparable of TMT-DDA in their ability to detect target engagement of losmapimod with MAPK14 and one of its downstream targets, MAPKAPK3. Using DIA for thermal shift quantitation is a cost-effective alternative to labeled quantitation in the TPP pipeline.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Proteome
/
Mitogen-Activated Protein Kinase 14
Type of study:
Diagnostic_studies
Language:
En
Journal:
J Proteome Res
Journal subject:
BIOQUIMICA
Year:
2023
Document type:
Article
Affiliation country:
United kingdom