PXD040212 is an
original dataset announced via ProteomeXchange.
Dataset Summary
Title | Mapping the Binding Interactions Between human Gasdermin D and human Caspase-1 Using Carbene Footprinting |
Description | Carbene footprinting is a recently developed mass spectrometry-based chemical labeling technique that probes protein interactions and conformation. Here, we use the methodology to investigate binding interactions between the protease human Caspase-1 (C285A) and full-length human Gasdermin D (hGSDMD), which are important in inflammatory cell death. GSDMD is cleaved by Caspase-1, releasing its N-terminal domain which oligomerizes in the membrane to form large pores, resulting in lytic cell death. Regions of reduced carbene labeling (masking), caused by protein binding, were observed for each partner in the presence of the other and were consistent with hCaspase-1 exosite and active-site interactions. Most notably, the results showed direct occupancy of hCaspase-1 active site by hGSDMD for the first time. Differential carbene labeling of full-length hGSDMD and the pore-forming N-terminal domain assembled in liposomes showed masking of the latter consistent with oligomeric assembly and insertion into the lipid bilayer. Interactions between Caspase-1 and the specific inhibitor VRT-043198 were also studied by this approach. In wild-type Caspase-1, VRT-043198 alkylates the active-site C285. Here we showed by carbene labeling that this inhibitor can occupy the active-site of a C285A mutant reversibly. These findings add considerably to our knowledge of the hCaspase-1-hGSDMD system. |
HostingRepository | PRIDE |
AnnounceDate | 2023-08-14 |
AnnouncementXML | Submission_2023-08-14_07:23:53.897.xml |
DigitalObjectIdentifier | |
ReviewLevel | Peer-reviewed dataset |
DatasetOrigin | Original dataset |
RepositorySupport | Unsupported dataset by repository |
PrimarySubmitter | JamesLloyd |
SpeciesList | scientific name: Homo sapiens (Human); NCBI TaxID: 9606; |
ModificationList | iodoacetamide derivatized residue |
Instrument | LTQ FT |
Dataset History
Revision | Datetime | Status | ChangeLog Entry |
0 | 2023-02-16 14:57:05 | ID requested | |
⏵ 1 | 2023-08-14 07:23:54 | announced | |
2 | 2023-11-14 06:52:27 | announced | 2023-11-14: Updated project metadata. |
Publication List
Keyword List
submitter keyword: GSDMD,Footprinting,LC-MS,Interaction,Caspase-1 |
Contact List
NeilOldham |
contact affiliation | Professor of Biomolecular Mass Spectrometry, Faculty of Science, Room A32 Chemistry, University Park, Nottingham, NG7 2RD, United Kingdom |
contact email | neil.oldham@nottingham.ac.uk |
lab head | |
JamesLloyd |
contact affiliation | University of Nottingham |
contact email | james.lloyd@nottingham.ac.uk |
dataset submitter | |
Full Dataset Link List
Dataset FTP location
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PRIDE project URI |
Repository Record List
[ + ]
[ - ]
- PRIDE
- PXD040212
- Label: PRIDE project
- Name: Mapping the Binding Interactions Between human Gasdermin D and human Caspase-1 Using Carbene Footprinting