Organelle Membrane Proteomics Reveals Differential Influence of Mycobacterial Lipoglycans on Macrophage Phagosome Maturation and Autophagosome Accumulation

The mycobacterial cell wall component lipoarabinomannan (LAM) has been described as one of the key virulence factors of <i>Mycobacterium tuberculosis</i>. Modification of the terminal arabinan residues of this lipoglycan with mannose caps in <i>M. tuberculosis</i> or with phosphoinositol caps in <i>Mycobacterium smegmatis</i> results in distinct host immune responses. Given that <i>M. tuberculosis</i> typically persists in the phagosomal vacuole after being phagocytosed by macrophages, we performed a proteomic analysis of that organelle after treatment of macrophages with LAMs purified from the two mycobacterial species. The quantitative changes in phagosomal proteins suggested a distinct role for mannose-capped LAM in modulating protein trafficking pathways that contribute to the arrest of phagosome maturation. Enlightened by our proteomic data, we performed further experiments to show that only the LAM from <i>M. tuberculosis</i> inhibits accumulation of autophagic vacuoles in the macrophage, suggesting a new function for this virulence-associated lipid