Mycobacterium tuberculosis Infection Manipulates the Glycosylation Machinery and the N-Glycoproteome of Human Macrophages and Their Microparticles

J Proteome Res. 2017 Jan 6;16(1):247-263. doi: 10.1021/acs.jproteome.6b00685. Epub 2016 Nov 4.


Tuberculosis (TB) remains a prevalent and lethal infectious disease. The glycobiology associated with Mycobacterium tuberculosis infection of frontline alveolar macrophages is still unresolved. Herein, we investigated the regulation of protein N-glycosylation in human macrophages and their secreted microparticles (MPs) used for intercellular communication upon M. tb infection. LC-MS/MS-based proteomics and glycomics were performed to monitor the regulation of glycosylation enzymes and receptors and the N-glycome in in vitro-differentiated macrophages and in isolated MPs upon M. tb infection. Infection promoted a dramatic regulation of the macrophage proteome. Most notably, significant infection-dependent down-regulation (4-26 fold) of 11 lysosomal exoglycosidases, e.g., β-galactosidase, β-hexosaminidases and α-/β-mannosidases, was observed. Relative weak infection-driven transcriptional regulation of these exoglycosidases and a stronger augmentation of the extracellular hexosaminidase activity demonstrated that the lysosome-centric changes may originate predominantly from infection-induced secretion of the lysosomal content. The macrophages showed heterogeneous N-glycan profiles and displayed significant up-regulation of complex-type glycosylation and concomitant down-regulation of paucimannosylation upon infection. Complementary intact N-glycopeptide analysis supported a subcellular-specific manipulation of the glycosylation machinery and altered glycosylation patterns of lysosomal N-glycoproteins within infected macrophages. Interestingly, the corresponding macrophage-derived MPs displayed unique N-glycome and proteome signatures supporting a preferential packaging from plasma membranes. The MPs were devoid of infection-dependent N-glycosylation signatures, but interestingly displayed increased levels of the glyco-initiating oligosaccharyltransferase complex and associated α-glucosidases that correlated with increased formation, N-glycan precursor levels and N-glycan density of infected MPs. In conclusion, this system-wide study provides new insight into the host- and pathogen-driven N-glycoproteome manipulation of macrophages in TB.

Keywords: LC−MS/MS; Mycobacterium tuberculosis; N-glycosylation; exoglycosidase; glycomics; glycoprofiling; glycoproteome; macrophage; microparticle; proteomics; tuberculosis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Carbohydrate Sequence
  • Cell Line
  • Cell-Derived Microparticles / chemistry
  • Cell-Derived Microparticles / metabolism*
  • Cell-Derived Microparticles / microbiology
  • Endoplasmic Reticulum / chemistry
  • Endoplasmic Reticulum / metabolism
  • Gene Expression Regulation
  • Glycoproteins / genetics*
  • Glycoproteins / metabolism
  • Glycoside Hydrolases / genetics
  • Glycoside Hydrolases / metabolism
  • Glycosylation
  • Glycosyltransferases / genetics
  • Glycosyltransferases / metabolism
  • Golgi Apparatus / chemistry
  • Golgi Apparatus / metabolism
  • Host-Pathogen Interactions*
  • Humans
  • Lectins / genetics
  • Lectins / metabolism
  • Lysosomes / chemistry
  • Lysosomes / metabolism
  • Macrophages / chemistry
  • Macrophages / metabolism*
  • Macrophages / microbiology
  • Mannose / chemistry
  • Mannose / metabolism
  • Mycobacterium tuberculosis / growth & development*
  • Mycobacterium tuberculosis / pathogenicity
  • Proteome / genetics*
  • Proteome / metabolism
  • Sialic Acids / chemistry
  • Sialic Acids / metabolism
  • Signal Transduction


  • Glycoproteins
  • Lectins
  • Proteome
  • Sialic Acids
  • Glycosyltransferases
  • Glycoside Hydrolases
  • Mannose