Concatemer Assisted Stoichiometry Analysis (CASA): targeted mass spectrometry for protein quantification.

ABSTRACT: Large multi-protein machines are central to multiple biological processes. However, stoichiometric determination of protein complex subunits in their native states presents a significant challenge. This study addresses the limitations of current tools in accuracy and precision by introducing concatemer-assisted stoichiometry analysis (CASA). CASA leverages stable isotope-labeled concatemers and liquid chromatography parallel reaction monitoring mass spectrometry (LC-PRM-MS) to achieve robust quantification of proteins with sub-femtomole sensitivity. As a proof-of-concept, CASA was applied to study budding yeast kinetochores. Stoichiometries were determined for ex vivo reconstituted kinetochore components, including the canonical H3 nucleosomes, centromeric (Cse4^CENP-A) nu

SUBMITTER: Cai J 

PROVIDER: S-EPMC11291133 | biostudies-literature | 2024 Jul

REPOSITORIES: biostudies-literature