5I82

First Crystal Structure of E.coli Based Recombinant Diphtheria Toxin Mutant CRM197

Summary for 5I82

• Entry DOI: 10.2210/pdb5i82/pdb
• Related: 4AE0 4AE1
• Descriptor: Diphtheria toxin, SULFATE ION, GLYCEROL, ... (4 entities in total)
• Functional Keywords: diphtheria toxin, crm197, csgid, recombinant, structural genomics, center for structural genomics of infectious diseases, transferase
• Biological source: Corynebacterium diphtheriae
• Total number of polymer chains: 4
• Total formula weight: 234313.97

Authors

Minasov, G.,Shuvalova, L.,Mishra, R.P.N.,Goel, A.,Dubrovska, I.,Anderson, W.F.,Center for Structural Genomics of Infectious Diseases (CSGID) (deposition date: 2016-02-18, release date: 2016-03-02, Last modification date: 2024-11-13)

Primary citation

Mishra, R.P.N.,Yadav, R.S.P.,Jones, C.,Nocadello, S.,Minasov, G.,Shuvalova, L.A.,Anderson, W.F.,Goel, A.
Structural and immunological characterization of E. coli derived recombinant CRM 197 protein used as carrier in conjugate vaccines.
Biosci.Rep., 38:-, 2018

PubMed Abstract: It is established that the immunogenicity of polysaccharides is enhanced by coupling them to carrier proteins. Cross reacting material (CRM), a nontoxic variant of diphtheria toxin (DT) is widely used carrier protein for polysaccharide conjugate vaccines. Conventionally, CRM is isolated by fermentation of C7 (β) cultures, which often suffers from low yield. Recently, several recombinant approaches have been reported with robust processes and higher yields, which will improve the affordability of CRM-based vaccines. Vaccine manufacturers require detailed analytical information to ensure that the CRM meets quality standards and regulatory requirements. In the present manuscript we have described detailed structural characteristics of based recombinant CRM (rCRM) carrier protein. The crystal structure of the based rCRM was found to be identical with the reported crystal structure of the C7 CRM produced in C7 strain (Protein Data Bank (PDB) ID: 4EA0) The crystal structure of rCRM was determined at 2.3 Å resolution and structure was submitted to the PDB with accession number ID 5I82. This is the first report of a crystal structure of derived recombinant CRM carrier protein. Furthermore, the rCRM was conjugated to Vi polysaccharide to generate Typhoid conjugate vaccine (Vi-rCRM) and its immunogenicity was evaluated in Balb/C Mice. The Vi-rCRM conjugate vaccine was found to generate strong primary α-Vi antibody response and also showed a booster response after subsequent vaccination in mice. Overall data suggest that based recombinant CRM exhibits structural and immunological similarity with the C7 CRM and can be used as a carrier protein in conjugate vaccine development.

PubMed: 29875175
DOI: 10.1042/BSR20180238

Experimental method

X-RAY DIFFRACTION (2.35 Å)