肽聚糖的消化及可溶性片段分析
Digestion of Peptidoglycan and Analysis of Soluble Fragments.
作者信息
Schaub Ryan E, Dillard Joseph P
机构信息
Medical Microbiology & Immunology, University of Wisconsin-Madison, Madison, USA.
出版信息
Bio Protoc. 2017 Aug 5;7(15). doi: 10.21769/BioProtoc.2438.
Abstract
Peptidoglycan (murein) is a vital component of the cell wall of nearly all bacteria, composed of sugars linked by short peptides. This protocol describes the purification of macromolecular peptidoglycan from cultured bacteria and the analysis of enzyme-digested peptidoglycan fragments using high performance liquid chromatography (HPLC). Digested peptidoglycan fragments can be identified by mass spectrometry, or predicted by comparing retention times with other published chromatograms. The quantitative nature of this method allows for the measurement of changes to peptidoglycan composition between different species of bacteria, growth conditions, or mutations. This method can determine the overall architecture of peptidoglycan, such as peptide stem length, the extent of cross-linking, and modifications. Muropeptide analysis has been used to study the function of peptidoglycan-associated proteins and the mechanisms by which bacteria acquire antibiotic resistance.
摘要
肽聚糖（胞壁质）是几乎所有细菌细胞壁的重要组成部分，由通过短肽连接的糖类组成。本方案描述了从培养的细菌中纯化大分子肽聚糖以及使用高效液相色谱（HPLC）分析酶消化的肽聚糖片段的方法。消化后的肽聚糖片段可以通过质谱鉴定，或通过与其他已发表的色谱图比较保留时间来预测。该方法的定量性质允许测量不同细菌物种、生长条件或突变之间肽聚糖组成的变化。此方法可以确定肽聚糖的整体结构，如肽茎长度、交联程度和修饰情况。胞壁肽分析已被用于研究肽聚糖相关蛋白的功能以及细菌获得抗生素抗性的机制。
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