PXD040212-2

PXD040212 is an original dataset announced via ProteomeXchange.

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	2023-11-14
AnnouncementXML	Submission_2023-11-14_06:52:26.845.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	James Lloyd
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	iodoacetamide derivatized residue
Instrument	LTQ FT

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.

Dataset with its publication pending

submitter keyword: GSDMD,Footprinting,LC-MS,Interaction,Caspase-1

Neil Oldham
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
James Lloyd
contact affiliation	University of Nottingham
contact email	james.lloyd@nottingham.ac.uk
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD040212
     1. Label: PRIDE project
     2. Name: Mapping the Binding Interactions Between human Gasdermin D and human Caspase-1 Using Carbene Footprinting