The Secretome Landscape of Escherichia Coli O157:H7: Deciphering the Cell-Surface, Outer Membrane Vesicle and Extracellular Subproteomes

Ricardo Monteiro,Ingrid Chafsey,Valentin Ageorges,Sabine Leroy,Christophe Chambon,Michel Hébraud,Valérie Livrelli,Mariagrazia Pizza,Alfredo Pezzicoli,Mickaël Desvaux

Journal of Proteomics（2021）

Université Clermont Auvergne | INRAE | Université Clermont-Auvergne | GSK

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Abstract

Among diarrheagenic E. coli (DEC), enterohaemorrhagic E. coli (EHEC) are the most virulent anthropozoonotic agents. The ability of bacterial cells to functionally interact with their surrounding essentially relies on the secretion of different protein effectors. To experimentally determine the repertoire of extracytoproteins in E. coli O157:H7, a subproteomic analysis was performed not only considering the extracellular milieu but the cell surface and outer membrane vesicles. Following a secretome-based approach, the proteins trafficking from the interior to the exterior of the cell were depicted considering cognate protein transport systems and subcellular localisation. Label-free quantitative analysis of the proteosurfaceome, proteovesiculome and exoproteome from E. coli O157:H7 grown in three different nutrient media revealed differential protein expression profiles and allowed defining the core and variant subproteomes. Network analysis further revealed the higher abundance of some protein clusters in chemically defined medium over rich complex medium, especially related to some outer membrane proteins, ABC transport and Type III secretion systems. This first comprehensive study of the EHEC secretome unravels the profound influence of environmental conditions on the extracytoplasmic proteome, provides new insight in the physiology of E. coli O157:H7 and identifies potentially important molecular targets for the development of preventive strategies against EHEC/STEC.

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Key words

Shiga toxin-producing E. coli (STEC),Subproteomic analysis,Secretome,Outer membrane vesicle,Cell-surface proteins,Extracellular proteins

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【要点】：本研究首次全面揭示了E. coli O157:H7的分泌组，发现环境条件对胞外蛋白组的深刻影响，并识别了潜在的预防策略分子靶标。

【方法】：采用分泌组学方法，对E. coli O157:H7的胞外环境、细胞表面和外部膜泡中的蛋白进行了亚蛋白组分析。

【实验】：通过在三种不同营养培养基中培养E. coli O157:H7，进行无标记定量分析，比较了胞外表面蛋白组、膜泡蛋白组和分泌蛋白组的表达谱，并使用网络分析揭示了不同条件下的蛋白聚类变化。