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Structural and mechanistic insights into the bacterial amyloid secretion channel CsgG

机译：对细菌淀粉样蛋白分泌通道CsgG的结构和机制的见解

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摘要

Curli are functional amyloid fibres that constitute the major protein component of the extracellular matrix in pellicle biofilms formed by Bacteroidetes and Proteobacteria (predominantly of the a and γ classes). They provide a fitness advantage in pathogenic strains and induce a strong pro-inflammatory response during bacteraemia. Curli formation requires a dedicated protein secretion machinery comprising the outer membrane lipoprotein CsgG and two soluble accessory proteins, CsgE and CsgF. Here we report the X-ray structure of Escherichia coli CsgG in a non-lipidated, soluble form as well as in its native membrane-extracted conformation. CsgG forms an oligomeric transport complex composed of nine anticodon-binding-domain-like units that give rise to a 36-stranded β-barrel that traverses the bilayer and is connected to a cage-like vestibule in the periplasm. The transmembrane and periplasmic domains are separated by a 0.9-nm channel constriction composed of three stacked concentric phenylalanine, asparagine and tyrosine rings that may guide the extended polypeptide substrate through the secretion pore. The specificity factor CsgE forms a nonameric adaptor that binds and closes off the periplasmic face of the secretion channel, creating a 24,000 A pre-constriction chamber. Our structural, functional and electrophysiological analyses imply that CsgG is an ungated, non-selective protein secretion channel that is expected to employ a diffusion-based, entropy-driven transport mechanism.
机译：Curli是功能性淀粉样蛋白纤维，构成了由拟杆菌和变形杆菌（主要是a和γ类）形成的薄膜生物膜中细胞外基质的主要蛋白质成分。它们在致病菌株中具有健身优势，并在菌血症期间诱导强烈的促炎反应。卷曲的形成需要专用的蛋白质分泌机制，该机制包括外膜脂蛋白CsgG和两种可溶性辅助蛋白CsgE和CsgF。在这里，我们以非脂质，可溶形式以及其天然膜提取构象报告大肠杆菌CsgG的X射线结构。 CsgG形成由9个反密码子结合结构域样单元组成的寡聚转运复合物，该复合物形成一个36链的β-桶，该桶穿过双层并与周质中的笼状前庭相连。跨膜结构域和周质结构域被一个0.9 nm的通道缩窄分隔开，该通道缩窄由三个堆叠的同心苯丙氨酸，天冬酰胺和酪氨酸环组成，可引导延伸的多肽底物穿过分泌孔。特异性因子CsgE形成一个非异构的衔接子，该衔接子结合并封闭了分泌通道的周质面，形成了一个24,000 A的预收缩腔。我们的结构，功能和电生理分析表明，CsgG是无门的，非选择性的蛋白分泌通道，有望采用基于扩散的，熵驱动的转运机制。

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  • 来源
    《Nature》 |2014年第7530期|250-253|共4页
  • 作者

    Parveen Goyal; Petya V. Krasteva; Nani Van Gerven; Francesca Gubellini; Imke Van den Broeck; Anastassia Troupiotis-Tsaielaki; Wim Jonckheere; Gerard Pehau-Arnaudet; Jerome S. Pinkner; Matthew R. Chapman; Scott J. Hultgren; Stefan Howorka; Remi Fronzes; Han Remaut;

  • 作者单位

    Structural and Molecular Microbiology, Structural Biology Research Center, VIB, Pleinlaan 2,1050 Brussels, Belgium,Structural Biology Brussels, Vrije Universiteit Brussel, Pleinlaan 2, 1050 Brussels, Belgium;

    Unite G5 Biologie structurale de la secretion bactenenne, Institut Pasteur, 25-28 rue du Docteur Roux, 75015 Paris, France,UMR 3528, CNRS, Institut Pasteur, 25-28 rue du Docteur Roux, 75015 Paris France;

    Structural and Molecular Microbiology, Structural Biology Research Center, VIB, Pleinlaan 2,1050 Brussels, Belgium,Structural Biology Brussels, Vrije Universiteit Brussel, Pleinlaan 2, 1050 Brussels, Belgium;

    Unite G5 Biologie structurale de la secretion bactenenne, Institut Pasteur, 25-28 rue du Docteur Roux, 75015 Paris, France,UMR 3528, CNRS, Institut Pasteur, 25-28 rue du Docteur Roux, 75015 Paris France;

    Structural and Molecular Microbiology, Structural Biology Research Center, VIB, Pleinlaan 2,1050 Brussels, Belgium,Structural Biology Brussels, Vrije Universiteit Brussel, Pleinlaan 2, 1050 Brussels, Belgium;

    Structure et Fonction des Membranes Biologiques (SFMB), Universite Libre de Bruxelles, 1050 Brussels, Belgium;

    Structural and Molecular Microbiology, Structural Biology Research Center, VIB, Pleinlaan 2,1050 Brussels, Belgium,Structural Biology Brussels, Vrije Universiteit Brussel, Pleinlaan 2, 1050 Brussels, Belgium;

    UMR 3528, CNRS, Institut Pasteur, 25-28 rue du Docteur Roux, 75015 Paris France;

    Department of Molecular Microbiology and Microbial Pathogenesis, Washington University in Saint Louis School of Medicine, St Louis, Missouri 63110-1010, USA;

    Department of Molecular, Cellular and Developmental Biology, University of Michigan, Ann Arhor Michigan 48109-1048. USA;

    Department of Molecular Microbiology and Microbial Pathogenesis, Washington University in Saint Louis School of Medicine, St Louis, Missouri 63110-1010, USA;

    Department of Chemistry, Institute for Structural and Molecular Biology, University College London, London WC1H 0AJ, UK;

    Unite G5 Biologie structurale de la secretion bactenenne, Institut Pasteur, 25-28 rue du Docteur Roux, 75015 Paris, France,UMR 3528, CNRS, Institut Pasteur, 25-28 rue du Docteur Roux, 75015 Paris France;

    Structural and Molecular Microbiology, Structural Biology Research Center, VIB, Pleinlaan 2,1050 Brussels, Belgium,Structural Biology Brussels, Vrije Universiteit Brussel, Pleinlaan 2, 1050 Brussels, Belgium;

  • 收录信息 美国《科学引文索引》(SCI);美国《工程索引》(EI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
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