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dc.contributor.authorShcherbakova, L
dc.contributor.authorPardo, M
dc.contributor.authorRoumeliotis, T
dc.contributor.authorChoudhary, J
dc.coverage.spatialEngland
dc.date.accessioned2023-01-24T13:40:33Z
dc.date.available2023-01-24T13:40:33Z
dc.date.issued2021-01-01
dc.identifier.citationWellcome Open Research, 2021, 6 pp. 260 -en_US
dc.identifier.issn2398-502X
dc.identifier.urihttps://repository.icr.ac.uk/handle/internal/5663
dc.identifier.eissn2398-502X
dc.identifier.eissn2398-502X
dc.identifier.doi10.12688/wellcomeopenres.17160.1
dc.identifier.doi10.12688/wellcomeopenres.17160.1
dc.description.abstractBackground: Cross-linking mass spectrometry (XL-MS) is a powerful technology capable of yielding structural insights across the complex cellular protein interaction network. However, up to date most of the studies utilising XL-MS to characterise individual protein complexes' topology have been carried out on over-expressed or recombinant proteins, which might not accurately represent native cellular conditions. Methods: We performed XL-MS using MS-cleavable crosslinker disuccinimidyl sulfoxide (DSSO) after immunoprecipitation of endogenous BRG/Brahma-associated factors (BAF) complex and co-purifying proteins. Data are available via ProteomeXchange with identifier PXD027611. Results: Although we did not detect the expected enrichment of crosslinks within the BAF complex, we identified numerous crosslinks between three co-purifying proteins, namely Thrap3, Bclaf1 and Erh. Thrap3 and Bclaf1 are mostly disordered proteins for which no 3D structure is available. The XL data allowed us to map interaction surfaces on these proteins, which overlap with the non-disordered portions of both proteins. The identified XLs are in agreement with homology-modelled structures suggesting that the interaction surfaces are globular. Conclusions: Our data shows that MS-cleavable crosslinker DSSO can be used to characterise in detail the topology and interaction surfaces of endogenous protein complexes without the need for overexpression. We demonstrate that Bclaf1, Erh and Thrap3 interact closely with each other, suggesting they might form a novel complex, hereby referred to as BET complex. This data can be exploited for modelling protein-protein docking to characterise the three-dimensional structure of the complex. Endogenous XL-MS might be challenging due to crosslinker accessibility, protein complex abundance or isolation efficiency, and require further optimisation for some complexes like the BAF complex to detect a substantial number of crosslinks.
dc.formatElectronic-eCollection
dc.format.extent260 -
dc.languageeng
dc.language.isoengen_US
dc.publisherF1000 Research Ltden_US
dc.relation.ispartofWellcome Open Research
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/en_US
dc.subjectBclaf1
dc.subjectCLMS cross-linking mass spectrometry
dc.subjectDSSO
dc.subjectEndogenous complexes
dc.subjectErh
dc.subjectThrap3
dc.subjectXL-MS
dc.titleIdentifying and characterising Thrap3, Bclaf1 and Erh interactions using cross-linking mass spectrometry.en_US
dc.typeJournal Article
dcterms.dateAccepted2021-09-24
dc.date.updated2023-01-24T13:18:29Z
rioxxterms.versionVoRen_US
rioxxterms.versionofrecord10.12688/wellcomeopenres.17160.1en_US
rioxxterms.licenseref.startdate2021-01-01
rioxxterms.typeJournal Article/Reviewen_US
pubs.author-urlhttps://www.ncbi.nlm.nih.gov/pubmed/35865489
pubs.organisational-group/ICR
pubs.organisational-group/ICR/Primary Group
pubs.organisational-group/ICR/Primary Group/ICR Divisions
pubs.organisational-group/ICR/Primary Group/ICR Divisions/Cancer Biology
pubs.organisational-group/ICR/Primary Group/ICR Divisions/Cancer Biology/Functional Proteomics Group
pubs.organisational-group/ICR/Students
pubs.organisational-group/ICR/Students/PhD and MPhil
pubs.organisational-group/ICR/Students/PhD and MPhil/18/19 Starting Cohort
pubs.publication-statusPublished online
pubs.publisher-urlhttp://dx.doi.org/10.12688/wellcomeopenres.17160.1
pubs.volume6
icr.researchteamFunctional Proteomicsen_US
dc.contributor.icrauthorShcherbakova, Liudmila
dc.contributor.icrauthorPardo Calvo, Maria Mercedes
icr.provenanceDeposited by Dr Mercedes Pardo Calvo on 2023-01-24. Deposit type is initial. No. of files: 1. Files: Identifying and characterising Thrap3, Bclaf1 and Erh interactions using cross-linking mass spectrometry.pdf


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