Isolation and analysis of cell wall components from Streptococcus pneumoniae
Isolation and analysis of cell wall components from Streptococcus pneumoniae
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DOI：
10.1016/j.ab.2011.11.026
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发表时间：
2012-02-15
影响因子：
2.9
通讯作者：
Vollmer, Waldemar
中科院分区：
文献类型：
--
作者：
Bui, Nhat Khai;Eberhardt, Alice;Vollmer, Waldemar
致病菌肺炎链球菌的复杂和异质性细胞壁由肽聚糖和共价附着的壁壁壁酸组成。网状的肽聚糖是由短肽交联的聚糖链形成的。我们已经开发了一种纯化聚糖链的方法，我们发现它们比大约25个双糖单位长。从纯化的肽聚糖中，我们释放了50种多肽，这些多肽的长度（三肽、四肽或五肽，有或没有单肽或二肽分支）、多肽交联程度（单体、二聚体或三聚体）、聚糖链上的修饰（n -去乙酰化、o-乙酰化，或缺乏GlcNAc或GlcNAc- murnac）或多肽（谷氨酸而不是谷氨酰胺）不同。我们还建立了一种分离壁壁酸链的方法，发现最丰富的链有6或7个重复单元。最后，我们获得了整个不溶性细胞壁的固态核磁共振谱。这些新工具将有助于表征突变菌株，细胞壁修饰酶和蛋白质-细胞壁相互作用。(C) 2011爱思唯尔公司版权所有。
The complex and heterogeneous cell wall of the pathogenic bacterium Streptococcus pneumoniae is composed of peptidoglycan and a covalently attached wall teichoic acid. The net-like peptidoglycan is formed by glycan chains that are crosslinked by short peptides. We have developed a method to purify the glycan chains, and we show that they are longer than approximately 25 disaccharide units. From purified peptidoglycan, we released 50 muropeptides that differ in the length of their peptides (tri-, tetra-, or pentapeptides with or without mono- or dipeptide branch), the degree of peptide crosslinking (monomer, dimer, or trimer), and the presence of modifications in the glycan chains (N-deacetylation, O-acetylation, or lack of GlcNAc or GlcNAc-MurNAc) or peptides (glutamic acid instead of glutamine). We also established a method to isolate wall teichoic acid chains and show that the most abundant chains have 6 or 7 repeating units. Finally, we obtained solid-state nuclear magnetic resonance spectra of whole insoluble cell walls. These novel tools will help to characterize mutant strains, cell wall-modifying enzymes, and protein-cell wall interactions. (C) 2011 Elsevier Inc. All rights reserved.