Isobaric crosslinking mass spectrometry technology for studying conformational and structural changes in proteins and complexes.
Publication Title
Elife
Document Type
Article
Publication Date
11-14-2024
Keywords
washington; isb; Mass Spectrometry; Protein Conformation; Cross-Linking Reagents; RNA Polymerase II; Proteins; Multiprotein Complexes; Humans
Abstract
Dynamic conformational and structural changes in proteins and protein complexes play a central and ubiquitous role in the regulation of protein function, yet it is very challenging to study these changes, especially for large protein complexes, under physiological conditions. Here, we introduce a novel isobaric crosslinker, Qlinker, for studying conformational and structural changes in proteins and protein complexes using quantitative crosslinking mass spectrometry. Qlinkers are small and simple, amine-reactive molecules with an optimal extended distance of ~10 Å, which use MS2 reporter ions for relative quantification of Qlinker-modified peptides derived from different samples. We synthesized the 2-plex Q2linker and showed that the Q2linker can provide quantitative crosslinking data that pinpoints key conformational and structural changes in biosensors, binary and ternary complexes composed of the general transcription factors TBP, TFIIA, and TFIIB, and RNA polymerase II complexes.
Specialty/Research Institute
Institute for Systems Biology
DOI
10.7554/eLife.99809