Improving Identification of In-organello Protein-Protein Interactions Using an Affinity-enrichable, Isotopically Coded, and Mass Spectrometry-cleavable Chemical Crosslinker

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dc.contributor.authorMakepeace, Karl A.T.
dc.contributor.authorMohammed, Yassene
dc.contributor.authorRudashevskaya, Elena L.
dc.contributor.authorPetrotchenko, Evgeniy V.
dc.contributor.authorVögtle, F.-Nora
dc.contributor.authorMeisinger, Chris
dc.contributor.authorSickmann, Albert
dc.contributor.authorBorchers, Christoph H.
dc.date.accessioned2021-01-26T18:15:48Z
dc.date.available2021-01-26T18:15:48Z
dc.date.copyright2020en_US
dc.date.issued2020
dc.description.abstractAn experimental and computational approach for identification of protein-protein interactions by ex vivo chemical crosslinking and mass spectrometry (CLMS) has been developed that takes advantage of the specific characteristics of cyanurbiotindipropionylsuccinimide (CBDPS), an affinity-tagged isotopically coded mass spectrometry (MS)-cleavable crosslinking reagent. Utilizing this reagent in combination with a crosslinker-specific data-dependent acquisition strategy based on MS2 scans, and a software pipeline designed for integrating crosslinker-specific mass spectral information led to demonstrated improvements in the application of the CLMS technique, in terms of the detection, acquisition, and identification of crosslinker-modified peptides. This approach was evaluated on intact yeast mitochondria, and the results showed that hundreds of unique protein-protein interactions could be identified on an organelle proteome-wide scale. Both known and previously unknown protein-protein interactions were identified. These interactions were assessed based on their known sub-compartmental localizations. Additionally, the identified crosslinking distance constraints are in good agreement with existing structural models of protein complexes involved in the mitochondrial electron transport chain.en_US
dc.description.reviewstatusRevieweden_US
dc.description.scholarlevelFacultyen_US
dc.description.sponsorshipThe University of Victoria-Genome BC Proteomics Centre is grateful for funding from Genome Canada and Genome BC for operations (204PRO) and technology development (214PRO) through the Genome Innovations Network, and for funding through the Genomics Technology Platform (264PRO). CHB would also like to thank the Natural Sciences and Engineering Research Council of Canada (NSERC) and the Leading Edge Endowment Fund for support. CHB is also grateful for support from the Segal McGill Chair in Molecular Oncology at McGill University (Montreal, Quebec, Canada), and for support from the Warren Y. Soper Charitable Trust and the Alvin Segal Family Foundation to the Jewish General Hospital (Montreal, Quebec, Canada). This work was also supported by the Deutsche Forschungsgemeinschaft (DFG) and the Excellence Initiative of the German Federal & State Governments (CIBSS - EXC-2189 - Project ID 390939984 and SFB1381 Project ID 403222702) to CM and FNV and the Emmy-Noether-Programm of the DFG to FNV.en_US
dc.identifier.citationMakepeace, K. A. T., Mohammed, Y., Rudashevskaya, E. L., Petrotchenko, E. V., Vögtle, F., Meisinger, C., … Borchers, C. H. (2020). Improving Identification of Inorganello Protein-Protein Interactions Using an Affinity-enrichable, Isotopically Coded, and Mass Spectrometry-cleavable Chemical Crosslinker. Molecular & Cellular Proteomics, 19(4), 624-639. https://doi.org/10.1074/mcp.RA119.001839.en_US
dc.identifier.urihttps://doi.org/10.1074/mcp.RA119.001839
dc.identifier.urihttp://hdl.handle.net/1828/12589
dc.language.isoenen_US
dc.publisherMolecular & Cellular Proteomicsen_US
dc.subjectUVic Genome BC Proteomics Centre
dc.subject.departmentDepartment of Biochemistry and Microbiology
dc.titleImproving Identification of In-organello Protein-Protein Interactions Using an Affinity-enrichable, Isotopically Coded, and Mass Spectrometry-cleavable Chemical Crosslinkeren_US
dc.typeArticleen_US

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