Towards an automated analysis of bacterial peptidoglycan structure.
Towards an automated analysis of bacterial peptidoglycan structure.
复制标题
DOI：
10.1007/s00216-016-9857-5
复制
发表时间：
2017-01
影响因子：
4.3
通讯作者：
Mesnage, Stephane
中科院分区：
文献类型：
--
作者：
Bern, Marshall;Beniston, Richard;Mesnage, Stephane
肽聚糖（PG）是细菌细胞被膜的重要组成部分。这种大分子由N-乙酰葡糖胺和N-乙酰胞壁酸交替的聚糖链组成，通过含有非标准氨基酸的短肽交联。PG的结构分析通常涉及聚糖链的酶促消化和通过反相HPLC分离二糖肽，然后收集MALDI-TOF和/或串联质谱法的单个峰。在这里，我们报告了一种新的策略，使用鸟枪蛋白质组学技术的系统和公正的结构分析PG使用高分辨率质谱和自动化分析HCD和ETD碎片光谱与Byonic软件。使用医院病原体艰难梭菌的PG作为概念的证明，我们表明，这种高通量方法允许识别先前描述的所有PG单体和二聚体，只留下3-3和4-3交联作为手动步骤的歧义。我们的分析证实了以前的研究结果，C。艰难肽聚糖主要包括脱乙酰化的N-乙酰葡糖胺残基和3-3个交联。该分析还揭示了许多低丰度的具有先前未报道的肽序列的胞肽。细菌细胞被膜包括质膜、肽聚糖和表面层。肽聚糖是细菌所特有的，也是最重要的抗生素的靶标;在这里，它是通过质谱分析的。本文的在线版本（doi：10.1007/s 00216 -016-9857-5）包含补充材料，可供授权用户使用。
Peptidoglycan (PG) is an essential component of the bacterial cell envelope. This macromolecule consists of glycan chains alternating N-acetylglucosamine and N-acetylmuramic acid, cross-linked by short peptides containing nonstandard amino acids. Structural analysis of PG usually involves enzymatic digestion of glycan strands and separation of disaccharide peptides by reversed-phase HPLC followed by collection of individual peaks for MALDI-TOF and/or tandem mass spectrometry. Here, we report a novel strategy using shotgun proteomics techniques for a systematic and unbiased structural analysis of PG using high-resolution mass spectrometry and automated analysis of HCD and ETD fragmentation spectra with the Byonic software. Using the PG of the nosocomial pathogen Clostridium difficile as a proof of concept, we show that this high-throughput approach allows the identification of all PG monomers and dimers previously described, leaving only disambiguation of 3–3 and 4–3 cross-linking as a manual step. Our analysis confirms previous findings that C. difficile peptidoglycans include mainly deacetylated N-acetylglucosamine residues and 3–3 cross-links. The analysis also revealed a number of low abundance muropeptides with peptide sequences not previously reported. The bacterial cell envelope includes plasma membrane, peptidoglycan, and surface layer. Peptidoglycan is unique to bacteria and the target of the most important antibiotics; here it is analyzed by mass spectrometry. The online version of this article (doi:10.1007/s00216-016-9857-5) contains supplementary material, which is available to authorized users.
登录
查看更多内容
影响因子：
4.6
作者：
Lambert C;Lerner TR;Bui NK;Somers H;Aizawa S;Liddell S;Clark A;Vollmer W;Lovering AL;Sockett RE
通讯作者：
Sockett RE
影响因子：
6.7
作者：
Frirdich E;Biboy J;Adams C;Lee J;Ellermeier J;Gielda LD;Dirita VJ;Girardin SE;Vollmer W;Gaynor EC
通讯作者：
Gaynor EC
影响因子：
4.8
作者：
Peltier, Johann;Courtin, Pascal;Pons, Jean-Louis
通讯作者：
Pons, Jean-Louis
影响因子：
4.6
作者：
Kühner D;Stahl M;Demircioglu DD;Bertsche U
通讯作者：
Bertsche U
影响因子：
4.8
作者：
Mainardi, JL;Fourgeaud, M;Arthur, M
通讯作者：
Arthur, M