Time-Resolved Study of In Situ Generated Protein Subcomplexes by Tandem-Trapped Ion Mobility Spectrometry.

ABSTRACT: We present an advanced analytical strategy that exploits the unique capabilities of tandem-trapped ion mobility spectrometry (Tandem-TIMS) to combine high-resolution ion mobility separation with targeted collisional activation and controlled gas-phase trapping. This approach enables the in situ generation of subcomplexes from native-like protein complexes of a selected charge state and allows direct evaluation of their kinetic stability and structural integrity in the gas phase. Using this workflow, we investigated the gas-phase stability of in situ generated streptavidin subunits. Streptavidin tetramers 15+ were mobility-selected and subjected to CID at 160 V between TIMS-1 and TIMS-2, producing monomers, dimers, and trimers. The resulting subcomplexes were then stored in TIMS-2 for up to

SUBMITTER: Pedrete T 

PROVIDER: S-EPMC12944212 | biostudies-literature | 2026 Feb

REPOSITORIES: biostudies-literature