Automating data analysis for hydrogen/deuterium exchange mass spectrometry using data-independent acquisition methodology.
Nat Commun
; 15(1): 2200, 2024 Mar 11.
Article
em En
| MEDLINE
| ID: mdl-38467655
ABSTRACT
We present a hydrogen/deuterium exchange workflow coupled to tandem mass spectrometry (HX-MS2) that supports the acquisition of peptide fragment ions alongside their peptide precursors. The approach enables true auto-curation of HX data by mining a rich set of deuterated fragments, generated by collisional-induced dissociation (CID), to simultaneously confirm the peptide ID and authenticate MS1-based deuteration calculations. The high redundancy provided by the fragments supports a confidence assessment of deuterium calculations using a combinatorial strategy. The approach requires data-independent acquisition (DIA) methods that are available on most MS platforms, making the switch to HX-MS2 straightforward. Importantly, we find that HX-DIA enables a proteomics-grade approach and wide-spread applications. Considerable time is saved through auto-curation and complex samples can now be characterized and at higher throughput. We illustrate these advantages in a drug binding analysis of the ultra-large protein kinase DNA-PKcs, isolated directly from mammalian cells.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Medição da Troca de Deutério
/
Hidrogênio
Limite:
Animals
Idioma:
En
Ano de publicação:
2024
Tipo de documento:
Article