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PXD020014

PXD020014 is an original dataset announced via ProteomeXchange.

Dataset Summary
Title"In-gel cross-linking mass spectrometry (IGX-MS) of proteins and protein complexes"
DescriptionCross-linking mass spectrometry has developed into an important method to study protein structures and interactions. The in-solution cross-linking workflows involve time and sample consuming steps and do not provide sensible solutions for differentiating cross-links obtained from co-occurring protein oligomers, complexes, or conformers. Here we developed a cross-linking workflow combining blue native PAGE with in-gel cross-linking mass spectrometry (IGX-MS). This workflow circumvents steps, such as buffer exchange and cross-linker concentration optimization. Additionally, IGX-MS enables the parallel analysis of co-occurring protein complexes using only small amounts of sample. Another benefit of IGX-MS observed by experiments on GroEL and purified bovine heart mitochondria, is the substantial reduction of artificial over-length cross-links when compared to in-solution cross-linking. We next used IGX-MS to investigate the complement components C5, C6, and their hetero-dimeric C5b6 complex. The obtained cross-links were used to generate a refined structural model of the complement component C6, resembling C6 in its inactivated state. This finding shows that IGX-MS can be used to provide new insights into the initial stages of the terminal complement pathway.
HostingRepositoryPRIDE
AnnounceDate2024-10-22
AnnouncementXMLSubmission_2024-10-22_05:16:41.787.xml
DigitalObjectIdentifier
ReviewLevelPeer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportUnsupported dataset by repository
PrimarySubmitterJohannes Hevler
SpeciesList scientific name: Escherichia coli; NCBI TaxID: 562; scientific name: Bos taurus (Bovine); NCBI TaxID: 9913; scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationListcarbamoylated residue; monohydroxylated residue; acetylated residue
InstrumentOrbitrap Fusion Lumos; Orbitrap Fusion
Dataset History
RevisionDatetimeStatusChangeLog Entry
02020-06-25 05:26:44ID requested
12020-12-14 05:22:18announced
22022-02-13 16:42:58announced2022-02-14: Updated project metadata.
32024-10-22 05:16:42announced2024-10-22: Updated project metadata.
Publication List
Dataset with its publication pending
Keyword List
ProteomeXchange project tag: EPIC-XS
submitter keyword: BN-PAGE/ Cross-linking/ Protein complexes/ Complement/ Protein modeling
Contact List
Albert J. R Heck
contact affiliationUtrecht University
contact emailA.J.R.Heck@uu.nl
lab head
Johannes Hevler
contact affiliationUtrecht University
contact emailj.f.hevler@uu.nl
dataset submitter
Full Dataset Link List
Dataset FTP location
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PRIDE project URI
Repository Record List
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