{"database":"biostudies-literature","file_versions":[],"scores":null,"additional":{"omics_type":["Unknown"],"submitter":["Cai J"],"funding":["NIH HHS","NIGMS NIH HHS"],"pubmed_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 <i>ex vivo</i> reconstituted kinetochore components, including the canonical H3 nucleosomes, centromeric (Cse4<sup>CENP-A</sup>) nu"],"journal":["bioRxiv : the preprint server for biology"],"pagination":["2024.07.26.605382"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC11291133"],"repository":["biostudies-literature"],"pubmed_title":["Concatemer Assisted Stoichiometry Analysis (CASA): targeted mass spectrometry for protein quantification."],"pmcid":["PMC11291133"],"funding_grant_id":["S10 OD023498","R01 GM116897","R01 GM151191"],"pubmed_authors":["Suhandynata RT","Cai J","Wang Z","Zhou H","Hinshaw SM","Yun Q","Zhang CY"],"additional_accession":[]},"is_claimable":false,"name":"Concatemer Assisted Stoichiometry Analysis (CASA): targeted mass spectrometry for protein quantification.","description":"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 <i>ex vivo</i> reconstituted kinetochore components, including the canonical H3 nucleosomes, centromeric (Cse4<sup>CENP-A</sup>) nu","dates":{"release":"2024-01-01T00:00:00Z","publication":"2024 Jul","modification":"2025-04-04T10:13:21.003Z","creation":"2025-04-04T10:13:21.003Z"},"accession":"S-EPMC11291133","cross_references":{"pubmed":["39091769"],"doi":["10.1101/2024.07.26.605382"]}}