Tyrosine-Reactive Cross-Linker for Probing Protein Three-Dimensional Structures

Cross-linking mass spectrometry (XL-MS) has made significant progress in understanding the structure of protein and elucidating architectures of larger protein complexes. Current XL-MS applications are limited to targeting lysine, glutamic acid, aspartic acid, and cysteine residues. There remains a need for the development of novel cross-linkers enabling selective targeting of other amino acid residues in proteins. Here, a novel simple crosslinker, namely, [4,4'-(disulfanediylbis(ethane-2,1-diyl)) bis(1,2,4-triazolidine-3,5-dione)] (DBB), has been designed, synthesized, and characterized. This cross-linker can react selectively with tyrosine residues in protein through the electrochemical click reaction. The DBB cross-links produced the characteristic peptides before and after electrochemical reduction, thus permitting the simplified data analysis and accurate identification for the cross-linked products. This is the first time a cross-linker is developed for targeting tyrosine residues on protein without using photoirradiation or a metal catalyst. This strategy might potentially be used as a complementary tool for XL-MS to probe protein 3D structures, protein complexes, and protein-protein interaction.

Automated summary

XL-MS has advanced in understanding protein structures and larger protein complexes by introducing a novel cross-linker, DBB, which can selectively target tyrosine residues in proteins through an electrochemical click reaction. This cross-linking strategy eliminates the need for photoirradiation or a metal catalyst, providing a potential complementary tool for XL-MS in probing protein 3D structures and interactions.