Biochemistry

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    Protocols in Current Issue
    Purification and HPLC Analysis of Cell Wall Muropeptides from Caulobacter crescentus
    Authors:  Gabriele Stankeviciute and Eric A. Klein, date: 11/05/2019, view: 3538, Q&A: 0
    [Abstract] The peptidoglycan sacculus, or cell wall, is what defines bacterial cell shape. Cell wall composition can be best characterized at the molecular level by digesting the peptidoglycan murein polymer into its muropeptide subunits and quantifying the abundance of muropeptides using high-pressure liquid chromatography. Certain features of the cell wall ...
    Bacterial Cell Wall Precursor Phosphatase Assays Using Thin-layer Chromatography (TLC) and High Pressure Liquid Chromatography (HPLC)
    Authors:  Manuel Pazos, Christian Otten and Waldemar Vollmer, date: 03/20/2018, view: 5522, Q&A: 0
    [Abstract] Peptidoglycan encases the bacterial cytoplasmic membrane to protect the cell from lysis due to the turgor. The final steps of peptidoglycan synthesis require a membrane-anchored substrate called lipid II, in which the peptidoglycan subunit is linked to the carrier lipid undecaprenol via a pyrophosphate moiety. Lipid II is the target of ...
    Analysis of N-acetylmuramic acid-6-phosphate (MurNAc-6P) Accumulation by HPLC-MS
    Authors:  Marina Borisova and Christoph Mayer, date: 08/05/2017, view: 7506, Q&A: 0
    [Abstract] We describe here in detail a high-performance liquid chromatography-mass spectrometry (HPLC-MS)-based method to determine N-acetylmuramic acid-6-phosphate (MurNAc-6P) in bacterial cell extracts. The method can be applied to both Gram-negative and Gram-positive bacteria, and as an example we use Escherichia coli cells in this ...
    Digestion of Peptidoglycan and Analysis of Soluble Fragments
    Authors:  Ryan E. Schaub and Joseph P. Dillard, date: 08/05/2017, view: 10710, Q&A: 0
    [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 ...



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