Reagents for Crosslinking Mass Spectrometry
JHU Ref #: C17605
Value Proposition:
• MS-cleavable crosslinkers for capturing protein-protein interactions.
• Compounds feature an NHS group and diazirine for covalent non-specific residue linkages.
• Avoid the N2 problem.
• Compounds are water-soluble and cell-permeable.
Technology Description
Researchers at Johns Hopkins have developed MS-cleavable heterobifunctional photo-crosslinkers that capture proteome-wide protein-protein interactions at residue-level resolution. The heterobifunctional probes comprise of a diazirine and N-hydroxy-succinimidyl (NHS) carbamate groups that crosslink interacting proteins. Additionally, these compounds are water-soluble and cell permeable allowing for use in both extracts and in cells.
Unmet Need
Crosslinking mass spectrometry (XL-MS) is a useful method at the crossroads of structural and cellular biology to interrogate protein-protein interactions at a proteome-wide scale. Early crosslinkers applied in XL-MS studies used homobifunctional compounds reliant on surface lysine residues. Photoactivated crosslinkers provide an alternative for non-specific residue crosslinking, but are complicated by the N2 problem where the search space associated with possible crosslinked peptides becomes exponentially complex. Therefore, there is a strong need for the development of next generation crosslinking compounds that allow for proteome-wide interrogation of protein-protein interactions and bypass the N2.
Stage of Development
Compounds synthesized and validated.
Data Availability
Data can be found at below publication.
Publication
Faustino, A, et al. Proteome-Wide Photo-Crosslinking Enables Residue-Level Visualization of Protein Interaction Networks in vivo. BioRxiv. 2022 Sept 20.
PCT WO2025064657 A1 patent application - Mass spectrometry-cleavable photo-crosslinkers for proteome-wide photo-crosslinking mass spectrometry
