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PXD057388

PXD057388 is an original dataset announced via ProteomeXchange.

Dataset Summary
TitleComparative analysis of the structural dynamics of diphtheria toxin and CRM197 carrier proteins used in the development of conjugate vaccines
DescriptionCarrier proteins are chemically linked to poorly immunogenic antigens to generate conjugate vaccines, which have significantly improved immunization strategies in recent decades. CRM197, a genetically detoxified diphtheria toxin (DT) mutant carrying the G52E mutation, is a widely used carrier protein as it retains lysine residues for antigen conjugation. In the past, CRM197 has been expressed in Corynebacterium diphtheriae, but low yields and high costs have prompted the exploration of alternative expression systems. Although high-yield expression and native refolding of CRM197 in E. coli are challenging due to its reducing cytoplasm, recent advances have enabled the production of soluble and well-folded recombinant CRM197 proteins, namely EcoCRM® and EcoCRM®(-Met). In this study, we use Hydrogen/Deuterium eXchange Mass Spectrometry (HDX-MS) to compare the structural dynamics of EcoCRM® and EcoCRM®(-Met) with DT wild-type. Our HDX-MS data show that the presence of the N-terminal methionine does not affect the structural dynamics of the two recombinant EcoCRM proteins. Furthermore, our results elucidate the molecular mechanism underlying the lack of toxicity of EcoCRM compared to DT wild-type: the G52E mutation in the CRM197 proteins exclusively alters the stability of the NAD-binding pocket and induces allosteric effects within the receptor-binding domain. Altogether, these insights support the substitution of CRM197 derived from Corynebacterium with the recombinant EcoCRM® and EcoCRM®(-Met) produced in E. coli, offering a cost-effective solution for use in conjugate vaccines.
HostingRepositoryPRIDE
AnnounceDate2025-04-07
AnnouncementXMLSubmission_2025-04-07_03:53:57.941.xml
DigitalObjectIdentifier
ReviewLevelPeer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportUnsupported dataset by repository
PrimarySubmitterSébastien Brier
SpeciesList scientific name: Escherichia coli; NCBI TaxID: 562; scientific name: Corynebacterium diphtheriae; NCBI TaxID: 1717;
ModificationListNo PTMs are included in the dataset
InstrumentSYNAPT G2-Si
Dataset History
RevisionDatetimeStatusChangeLog Entry
02024-10-31 03:40:05ID requested
12025-04-07 03:53:58announced
Publication List
10.1016/j.ijpharm.2025.125535;
Briday M, Carvalho N, Oganesyan N, Chang MJ, Lees A, Brier S, Chenal A, carrier proteins used in the development of conjugate vaccines. Int J Pharm, 675():125535(2025) [pubmed]
Keyword List
ProteomeXchange project tag: Hydrogen Deuterium Exchange (HDX-MS)
submitter keyword: comparative analysis, CRM197, structural characterization,Hydrogen/Deuterium eXchange mass spectrometry (HDX-MS), Biopharmaceutical products
Contact List
Sébastien Brier
contact affiliationInstitut Pasteur, Université Paris Cité, BioNMR and HDX-MS facility, Centre for Technological Resources and Research, Chemistry and Structural Biology Department, UMR CNRS 3528, F-75015 Paris, France
contact emailsebastien.brier@pasteur.fr
lab head
Sébastien Brier
contact affiliationInstitut Pasteur, Université Paris Cité, BioNMR and HDX-MS facility, Center for Technological Resources and Research, Chemistry and Structural Biology Department, UMR CNRS 3528, F-75015 Paris, France
contact emailsebastien.brier@pasteur.fr
dataset submitter
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