7RRW

Monomeric CRM197 expressed in E. coli

7RRW の概要

• エントリーDOI: 10.2210/pdb7rrw/pdb
• 分子名称: Diphtheria toxin, 2-AMINO-2-HYDROXYMETHYL-PROPANE-1,3-DIOL (3 entities in total)
• 機能のキーワード: inactivated diphtheria toxin, vaccine carrier, e. coli expression, toxin
• 由来する生物種: Corynebacterium diphtheriae
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 58605.56

構造登録者

Gallagher, D.T.,Lees, A. (登録日: 2021-08-10, 公開日: 2022-11-09, 最終更新日: 2024-10-23)

主引用文献

Gallagher, D.T.,Oganesyan, N.,Lees, A.
Monomeric crystal structure of the vaccine carrier protein CRM 197 and implications for vaccine development.
Acta Crystallogr.,Sect.F, 79:82-86, 2023

PubMed Abstract: CRM is a genetically detoxified mutant of diphtheria toxin (DT) that is widely used as a carrier protein in conjugate vaccines. Protective immune responses to several bacterial diseases are obtained by coupling CRM to glycans from these pathogens. Wild-type DT has been described in two oligomeric forms: a monomer and a domain-swapped dimer. Their proportions depend on the chemical conditions and especially the pH, with a large kinetic barrier to interconversion. A similar situation occurs in CRM, where the monomer is preferred for vaccine synthesis. Despite 30 years of research and the increasing application of CRM in conjugate vaccines, until now all of its available crystal structures have been dimeric. Here, CRM was expressed as a soluble, intracellular protein in an Escherichia coli strain engineered to have an oxidative cytoplasm. The purified product, called EcoCRM, remained monomeric throughout crystallization. The structure of monomeric EcoCRM is reported at 2.0 Å resolution with the domain-swapping hinge loop (residues 379-387) in an extended, exposed conformation, similar to monomeric wild-type DT. The structure enables comparisons across expression systems and across oligomeric states, with implications for monomer-dimer interconversion and for the optimization of conjugation.

PubMed: 36995122
DOI: 10.1107/S2053230X23002364

実験手法

X-RAY DIFFRACTION (2 Å)