Transient Cross-linking Mass Spectrometry: Taking Conformational Snapshots of Proteins DOI Creative Commons
Yuxin Xie, Jiawen Wang, Lei Yang

et al.

Analytical Chemistry, Journal Year: 2025, Volume and Issue: unknown

Published: March 4, 2025

The dynamic nature of protein conformations is central to their biological functions. Conventional structural biology techniques provide static snapshots, whereas a comprehensive understanding requires an analysis the conformations. In this study, we develop transient cross-linking mass spectrometry method using photo-cross-linker DCD. This cross-linker can be transiently activated accomplish cross-linking, and with sample freezing, are preserved, allowing temporal control on-demand cross-linking. Its site covers all amino acids, exhibiting diversity providing rich information. Additionally, data-processing strategy by integrating DCD-specific reporter ion defined ambiguous annotation criterion, thereby ensuring confidence in identification cross-link annotation. Thus, developed spectrometry, leveraging distinctive features DCD, has enabled us analyze complexes high resolution, take conformational discern coexistence intermediates, decipher fluctuations, shedding light on how proteins conformationally respond signals engage interacting partners. Our results highlight DCD's potential for probing changes, facilitating elucidation pivotal roles within systems.

Language: Английский

Transient Cross-linking Mass Spectrometry: Taking Conformational Snapshots of Proteins DOI Creative Commons
Yuxin Xie, Jiawen Wang, Lei Yang

et al.

Analytical Chemistry, Journal Year: 2025, Volume and Issue: unknown

Published: March 4, 2025

The dynamic nature of protein conformations is central to their biological functions. Conventional structural biology techniques provide static snapshots, whereas a comprehensive understanding requires an analysis the conformations. In this study, we develop transient cross-linking mass spectrometry method using photo-cross-linker DCD. This cross-linker can be transiently activated accomplish cross-linking, and with sample freezing, are preserved, allowing temporal control on-demand cross-linking. Its site covers all amino acids, exhibiting diversity providing rich information. Additionally, data-processing strategy by integrating DCD-specific reporter ion defined ambiguous annotation criterion, thereby ensuring confidence in identification cross-link annotation. Thus, developed spectrometry, leveraging distinctive features DCD, has enabled us analyze complexes high resolution, take conformational discern coexistence intermediates, decipher fluctuations, shedding light on how proteins conformationally respond signals engage interacting partners. Our results highlight DCD's potential for probing changes, facilitating elucidation pivotal roles within systems.

Language: Английский

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