疫苗载体蛋白CRM197的单体晶体结构及其对疫苗开发的意义

IF 1.1 4区 生物学 Q4 BIOCHEMICAL RESEARCH METHODS
D. Travis Gallagher, Natalia Oganesyan, Andrew Lees
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引用次数: 1

摘要

CRM197是白喉毒素(DT)的一种基因解毒突变体,广泛用作结合疫苗中的载体蛋白。通过将CRM197与这些病原体的聚糖偶联,可以获得对几种细菌性疾病的保护性免疫反应。野生型DT有两种低聚形式:单体和结构域交换的二聚体。它们的比例取决于化学条件,尤其是pH,相互转化有很大的动力学障碍。类似的情况发生在CRM197中,其中单体优选用于疫苗合成。尽管经过30年的研究,CRM197在偶联疫苗中的应用越来越多,但到目前为止,其所有可用的晶体结构都是二聚体的。在这里,CRM197在被改造成具有氧化细胞质的大肠杆菌菌株中作为可溶性细胞内蛋白表达。被称为EcoCRM的纯化产物在整个结晶过程中保持单体。据报道,单体EcoCRM的结构为2.0 Å分辨率,具有延伸、暴露构象的结构域交换铰链环(残基379-387),类似于单体野生型DT。该结构能够在表达系统和寡聚态之间进行比较,对单体-二聚体的相互转化和偶联的优化具有重要意义。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Monomeric crystal structure of the vaccine carrier protein CRM197 and implications for vaccine development

Monomeric crystal structure of the vaccine carrier protein CRM197 and implications for vaccine development

CRM197 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 CRM197 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 CRM197, where the monomer is preferred for vaccine synthesis. Despite 30 years of research and the increasing application of CRM197 in conjugate vaccines, until now all of its available crystal structures have been dimeric. Here, CRM197 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.

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来源期刊
Acta crystallographica. Section F, Structural biology communications
Acta crystallographica. Section F, Structural biology communications BIOCHEMICAL RESEARCH METHODSBIOCHEMISTRY &-BIOCHEMISTRY & MOLECULAR BIOLOGY
CiteScore
1.90
自引率
0.00%
发文量
95
期刊介绍: Acta Crystallographica Section F is a rapid structural biology communications journal. Articles on any aspect of structural biology, including structures determined using high-throughput methods or from iterative studies such as those used in the pharmaceutical industry, are welcomed by the journal. The journal offers the option of open access, and all communications benefit from unlimited free use of colour illustrations and no page charges. Authors are encouraged to submit multimedia content for publication with their articles. Acta Cryst. F has a dedicated online tool called publBio that is designed to make the preparation and submission of articles easier for authors.
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