Purification and Analysis of Mycobacterial Phosphatidylinositol Mannosides, Lipomannan, and Lipoarabinomannan

  • Kathryn C. Rahlwes
  • Julia Puffal
  • Yasu S. MoritaEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1954)


Mycobacteria and related bacteria in the Actinobacteria phylum are unusual in that they produce phosphatidylinositol (PI) as a major phospholipid species. PI can be further modified by glycan polymers, leading to the synthesis of PI mannosides (PIMs), lipomannan (LM), and lipoarabinomannan (LAM). Small lipids such as PI and PIMs are extracted with a mixture of chloroform, methanol, and water and analyzed by thin layer chromatography. For larger glycolipids, such as LM and LAM, more hydrophilic solvent is needed for the extraction, and SDS-PAGE is better suited for the analysis. For LM, further structural characterization can be performed by MALDI-TOF mass spectrometry. Precise quantification of PIMs, LM, and LAM can be performed by quantification of glycan staining using analytical software. The metabolic radiolabeling protocol is also described.

Key words

Glycolipids Glycan staining Lipomannan Lipoarabinomannan Phosphatidylinositol mannosides 



This work was supported by a Biomedical Research Grant (RG-414805) from the American Lung Association and a Research Grant from the Pittsfield Anti-Tuberculosis Association to YSM.


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Kathryn C. Rahlwes
    • 1
  • Julia Puffal
    • 1
  • Yasu S. Morita
    • 1
    Email author
  1. 1.Department of MicrobiologyUniversity of MassachusettsAmherstUSA

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