Effects of Chemical Cross-Linking on the Structure of Proteins and Protein Assemblies.

ABSTRACT: Chemical cross-linking is frequently used in structural biology for protein stabilization and probing protein structures, often in combination with mass spectrometry (XL-MS). These applications assume that chemical cross-linking does not significantly perturb the protein structure. Here, we directly tested this assumption by monitoring the time course of small-angle X-ray scattering (SAXS) signals from cross-linked protein samples. We investigated two common cross-linking reagents, bis(sulfosuccinimidyl)suberate (BS³) and formaldehyde (FA), with several protein systems ranging from large microtubule filaments down to globular proteins. Across all the measured protein systems, the results consistently showed that BS³ did not significantly change the protein structures,

SUBMITTER: Tayri-Wilk T 

PROVIDER: S-EPMC12291049 | biostudies-literature | 2025 Jul

REPOSITORIES: biostudies-literature