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Structure, dynamics and immunogenicity of a catalytically inactive CXC chemokine-degrading protease SpyCEP from Streptococcus pyogenes.
Computational and Structural Biotechnology Journal ( IF 6 ) Pub Date : 2020-03-13 , DOI: 10.1016/j.csbj.2020.03.004
Sophie McKenna 1 , Enrico Malito 2 , Sarah L Rouse 1 , Francesca Abate 3 , Giuliano Bensi 4 , Emiliano Chiarot 4 , Francesca Micoli 5 , Francesca Mancini 5 , Danilo Gomes Moriel 5 , Guido Grandi 6 , Danuta Mossakowska 7 , Max Pearson 8 , Yingqi Xu 1 , James Pease 9 , Shiranee Sriskandan 8 , Immaculada Margarit 4 , Matthew J Bottomley 2 , Stephen Matthews 1
Affiliation  

Over 18 million disease cases and half a million deaths worldwide are estimated to be caused annually by Group A Streptococcus. A vaccine to prevent GAS disease is urgently needed. SpyCEP (Streptococcus pyogenes Cell-Envelope Proteinase) is a surface-exposed serine protease that inactivates chemokines, impairing neutrophil recruitment and bacterial clearance, and has shown promising immunogenicity in preclinical models. Although SpyCEP structure has been partially characterized, a more complete and higher resolution understanding of its antigenic features would be desirable prior to large scale manufacturing. To address these gaps and facilitate development of this globally important vaccine, we performed immunogenicity studies with a safety-engineered SpyCEP mutant, and comprehensively characterized its structure by combining X-ray crystallography, NMR spectroscopy and molecular dynamics simulations. We found that the catalytically-inactive SpyCEP antigen conferred protection similar to wild-type SpyCEP in a mouse infection model. Further, a new higher-resolution crystal structure of the inactive SpyCEP mutant provided new insights into this large chemokine protease comprising nine domains derived from two non-covalently linked fragments. NMR spectroscopy and molecular simulation analyses revealed conformational flexibility that is likely important for optimal substrate recognition and overall function. These combined immunogenicity and structural data demonstrate that the full-length SpyCEP inactive mutant is a strong candidate human vaccine antigen. These findings show how a multi-disciplinary study was used to overcome obstacles in the development of a GAS vaccine, an approach applicable to other future vaccine programs. Moreover, the information provided may also facilitate the structure-based discovery of small-molecule therapeutics targeting SpyCEP protease inhibition.



中文翻译:

来自化脓性链球菌的催化失活 CXC 趋化因子降解蛋白酶 SpyCEP 的结构、动力学和免疫原性。

据估计,全世界每年有超过 1800 万例疾病病例和 50 万人死亡是由 A 组链球菌引起的。迫切需要一种预防 GAS 疾病的疫苗。SpyCEP(化脓性链球菌细胞包膜蛋白酶)是一种表面暴露的丝氨酸蛋白酶,可灭活趋化因子,损害中性粒细胞募集和细菌清除,并在临床前模型中显示出有希望的免疫原性。尽管 SpyCEP 结构已被部分表征,但在大规模制造之前仍需要对其抗原特征进行更完整和更高分辨率的了解。为了解决这些差距并促进这种全球重要疫苗的开发,我们对安全工程的 SpyCEP 突变体进行了免疫原性研究,并通过结合 X 射线晶体学、核磁共振波谱和分子动力学模拟来全面表征其结构。我们发现,催化失活的 SpyCEP 抗原在小鼠感染模型中提供了与野生型 SpyCEP 类似的保护作用。此外,失活的 SpyCEP 突变体的新的更高分辨率晶体结构为这种大型趋化因子蛋白酶提供了新的见解,该趋化因子蛋白酶包含源自两个非共价连接片段的九个结构域。核磁共振波谱和分子模拟分析揭示了构象灵活性,这对于最佳底物识别和整体功能可能很重要。这些组合的免疫原性和结构数据表明全长 SpyCEP 无活性突变体是强有力的候选人类疫苗抗原。这些发现展示了如何利用多学科研究来克服 GAS 疫苗开发中的障碍,这种方法适用于其他未来的疫苗项目。此外,所提供的信息还可能有助于基于结构的发现针对 SpyCEP 蛋白酶抑制的小分子疗法。

更新日期:2020-03-13
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