Sum of the parts: composition and architecture of the bacterial extracellular matrix.
Sum of the parts: composition and architecture of the bacterial extracellular matrix.
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DOI：
10.1016/j.jmb.2013.06.022
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发表时间：
2013-11-15
影响因子：
5.6
通讯作者：
Cegelski, Lynette
中科院分区：
文献类型：
--
作者：
McCrate, Oscar A.;Zhou, Xiaoxue;Reichhardt, Courtney;Cegelski, Lynette
细菌生物膜是一种复杂的多细胞集合体，对抗生素具有耐药性，是严重和慢性传染病的致病因素。需要新的方法和定量数据来确定细菌生物膜的分子组成。众所周知，大肠杆菌生物膜含有多糖类和功能性淀粉样纤维，称为卷曲，但在分子水平上准确确定生物膜组成一直难以捉摸。确定细胞外基质组成的能力对于阐明结构-功能关系至关重要，这将有助于制定破坏生物膜的化学策略。我们开发了一种将细胞外基质的非扰动制备与电子显微镜、生物化学和固态核磁共振光谱相结合的方法，以确定一种临床上重要的致病细菌--尿路致病性大肠杆菌--完整和不溶的细胞外基质的化学成分。我们的数据允许所有部分的总和分析。电子显微镜显示，超分子壳状结构包裹着单个细胞，并包裹着细菌群落。对基质和组成部分的生化和固体核磁共振测量表明，基质由两个主要组分组成，即卷曲和纤维素，每个组分的数量都是可以量化的。这种量化基质成分的方法广泛适用于其他生物和检查生物膜抑制剂的影响。总而言之，我们的核磁共振谱和纯化的ECM的电子显微镜启发我们将生物膜基质视为具有特定组成和结构的聚合物的集合，而不是未定义的黏液。
Bacterial biofilms are complex multicellular assemblies that exhibit resistance to antibiotics and contribute to the pathogenesis of serious and chronic infectious diseases. New approaches and quantitative data are needed to define the molecular composition of bacterial biofilms. E. coli biofilms are known to contain polysaccharides and functional amyloid fibers termed curli, yet accurate determinations of biofilm composition at the molecular level have been elusive. The ability to define the composition of the extracellular matrix is crucial for the elucidation of structure–function relationships that will aid the development of chemical strategies to disrupt biofilms. We have developed an approach that integrates non-perturbative preparation of the extracellular matrix with electron microscopy, biochemistry and solid-state NMR spectroscopy to define the chemical composition of the intact and insoluble extracellular matrix of a clinically important pathogenic bacterium—uropathogenic E. coli. Our data permitted a sum-of-all-the-parts analysis. Electron microscopy revealed supramolecular shell-like structures that encapsulated single cells and enmeshed the bacterial community. Biochemical and solid-state NMR measurements of the matrix and constitutive parts established that the matrix is composed of two major components, curli and cellulose, each in a quantifiable amount. This approach to quantifying the matrix composition is widely applicable to other organisms and to examining the influence of biofilm inhibitors. Collectively, our NMR spectra and the electron micrographs of the purified ECM inspire us to consider the biofilm matrix not as an undefined slime, but as an assembly of polymers with a defined composition and architecture.
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影响因子：
6.7
作者：
Kai-Larsen Y;Lüthje P;Chromek M;Peters V;Wang X;Holm A;Kádas L;Hedlund KO;Johansson J;Chapman MR;Jacobson SH;Römling U;Agerberth B;Brauner A
通讯作者：
Brauner A
影响因子：
3.2
作者：
Da Re, S;Ghigo, JM
通讯作者：
Ghigo, JM
影响因子：
4.4
作者：
BENNETT, AE;RIENSTRA, CM;GRIFFIN, RG
通讯作者：
GRIFFIN, RG
影响因子：
4.4
作者：
Lim, Ji Youn;May, Janine M.;Cegelski, Lynette
通讯作者：
Cegelski, Lynette
影响因子：
14.8
作者：
Cegelski, Lynette;Pinkner, Jerome S.;Hammer, Neal D.;Cusumano, Corinne K.;Hung, Chia S.;Chorell, Erik;Aberg, Veronica;Walker, Jennifer N.;Seed, Patrick C.;Almqvist, Fredrik;Chapman, Matthew R.;Hultgren, Scott J.
通讯作者：
Hultgren, Scott J.