Year: 2025
Type: article
Abstract: Cross-linking mass spectrometry is rapidly becoming a choice method for determining a protein's higher-order structure as well as capturing inter-protein interactions. In particular, diazirene-based photo-activatable cross-linkers, such as sulfo-SDA have been shown to be effective at generating high-density cross-linking data. Previously, we have shown that this method may be used to study binding orientation between two non-covalently linked complexes; however, several unexpected ions were noted in the MS2 spectra. In this study, the tandem mass spectrometry fragmentation patterns of sulfo-SDA-initiated cross-linked peptides under higher-energy collision induced (HCD), collision induced (CID) and electron transfer with supplementary HCD (EThcD) dissociations are discussed. The analysis revealed substantial insights into localising cross-linking sites, which is essential for accurate determination of protein higher-order structural characteristics.
Authors: Thomas Powell, Martin Ebner, Andrew Creese
Institution: Immunocore (United Kingdom)
Language: English
Subfield: Spectroscopy
Field: Chemistry
Domain: Physical Sciences
Open Access status: hybrid
Estimated APC paid: $4,150
Funder: Immunocore
OpenAlex ID: W4410104415