PXD057388 is an
original dataset announced via ProteomeXchange.
Dataset Summary
Title | Comparative analysis of the structural dynamics of diphtheria toxin and CRM197 carrier proteins used in the development of conjugate vaccines |
Description | Carrier 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. |
HostingRepository | PRIDE |
AnnounceDate | 2025-04-07 |
AnnouncementXML | Submission_2025-04-07_03:53:57.941.xml |
DigitalObjectIdentifier | |
ReviewLevel | Peer-reviewed dataset |
DatasetOrigin | Original dataset |
RepositorySupport | Unsupported dataset by repository |
PrimarySubmitter | Sébastien Brier |
SpeciesList | scientific name: Escherichia coli; NCBI TaxID: 562; scientific name: Corynebacterium diphtheriae; NCBI TaxID: 1717; |
ModificationList | No PTMs are included in the dataset |
Instrument | SYNAPT G2-Si |
Dataset History
Revision | Datetime | Status | ChangeLog Entry |
0 | 2024-10-31 03:40:05 | ID requested | |
⏵ 1 | 2025-04-07 03:53:58 | announced | |
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 affiliation | Institut 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 email | sebastien.brier@pasteur.fr |
lab head | |
Sébastien Brier |
contact affiliation | Institut 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 email | sebastien.brier@pasteur.fr |
dataset submitter | |
Full Dataset Link List
Dataset FTP location
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PRIDE project URI |
Repository Record List
[ + ]
[ - ]
- PRIDE
- PXD057388
- Label: PRIDE project
- Name: Comparative analysis of the structural dynamics of diphtheria toxin and CRM197 carrier proteins used in the development of conjugate vaccines