PXD020014 is an
original dataset announced via ProteomeXchange.
Dataset Summary
Title | "In-gel cross-linking mass spectrometry (IGX-MS) of proteins and protein complexes" |
Description | Cross-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. |
HostingRepository | PRIDE |
AnnounceDate | 2024-10-22 |
AnnouncementXML | Submission_2024-10-22_05:16:41.787.xml |
DigitalObjectIdentifier | |
ReviewLevel | Peer-reviewed dataset |
DatasetOrigin | Original dataset |
RepositorySupport | Unsupported dataset by repository |
PrimarySubmitter | Johannes 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; |
ModificationList | carbamoylated residue; monohydroxylated residue; acetylated residue |
Instrument | Orbitrap Fusion Lumos; Orbitrap Fusion |
Dataset History
Revision | Datetime | Status | ChangeLog Entry |
0 | 2020-06-25 05:26:44 | ID requested | |
1 | 2020-12-14 05:22:18 | announced | |
2 | 2022-02-13 16:42:58 | announced | 2022-02-14: Updated project metadata. |
⏵ 3 | 2024-10-22 05:16:42 | announced | 2024-10-22: 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 affiliation | Utrecht University |
contact email | A.J.R.Heck@uu.nl |
lab head | |
Johannes Hevler |
contact affiliation | Utrecht University |
contact email | j.f.hevler@uu.nl |
dataset submitter | |
Full Dataset Link List
Dataset FTP location
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PRIDE project URI |
Repository Record List
[ + ]
[ - ]
- PRIDE
- PXD020014
- Label: PRIDE project
- Name: "In-gel cross-linking mass spectrometry (IGX-MS) of proteins and protein complexes"