Characterization of Peptidoglycan Trimers after Gel Chromatography and Reversed-phase High-performance Liquid Chromatography by Positive-ion Plasma Desorption Mass Spectrometry

✍ Scribed by Andrea Zenker; Ernst Pittenauer; Beatrix Pfanzagl; Wolfgang Löffelhardt; Günter Allmaier

• Publisher: John Wiley and Sons
• Year: 1996
• Tongue: English
• Weight: 504 KB
• Volume: 10
• Category: Article
• ISSN: 0951-4198
• DOI: 10.1002/(sici)1097-0231(199612)10:15<1956::aid-rcm780>3.0.co;2-d

✦ Synopsis

A strategy for the primary stucture characterization of reduced peptidoglycan trimers derived from muramidase-digested murein (e.g. isolated from the cyanelles of Cyunophoru puradoxu) is outlined. First, muropeptides are separated by gel filtration according to their size (degree of cross-linking). This step is followed by reduction with sodium borohydride and reversed-phase high-performance liquid chromatography (HPLC). Wmeric and oligomeric compounds (molecular weight range 2500-4500 Da), in particular, are present in small quantities and therefore sophisticated methods for characterization are required due to the biological importance of these components. For determining moecular weight with high accuracy, positive-ion plasma desorption mass spectrometry (PDMS) proves to be a well-suited analytical method with sufficient sensitivity (medium picomole range) and mass accuracy (~0.04%). Based on combined data from PDMS, gel chromatography, HPLC and amino acid and amino sugar analyses, the primary structure of peptidoglycan trimeric compounds could be determined unambiguously.