Dataset Information

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

ABSTRACT: 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.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Macrophage

DISEASE(S): Tuberculosis

SUBMITTER: Ling Lee

LAB HEAD: Morten Thaysen-Andersen

PROVIDER: PXD005169 | Pride | 2024-01-26

REPOSITORIES: Pride

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			Action	DRS
	150404_ly_1B_1 (UI1 F033035).mzid_150404_ly_1B_1_(UI1_F033035).pride.mgf.gz	Mzid
	150404_ly_1B_1 (UI1 F033035).pride.mztab.gz	Mztab
	150404_ly_1B_1__UI1_F033035_.mzid.gz	Mzid
	150404_ly_1B_1__UI1_F033035_.mzid_150404_ly_1B_1__UI1_F033035_.MGF	Mzid
	150404_ly_2B_1 (UI2 F033043).mzid_150404_ly_2B_1_(UI2_F033043).pride.mgf.gz	Mzid

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Publications

Profound <i>N</i>-glycan remodelling accompanies MHC-II immunopeptide presentation.

Goodson Hayley H Kawahara Rebeca R Chatterjee Sayantani S Goncalves Gabriel G Fehring Joshua J Purcell Anthony W AW Croft Nathan P NP Thaysen-Andersen Morten M

Frontiers in immunology 20231108

Immunopeptidomics, the study of peptide antigens presented on the cell surface by the major histocompatibility complex (MHC), offers insights into how our immune system recognises self/non-self in health and disease. We recently discovered that hyper-processed (remodelled) <i>N</i>-glycans are dominant features decorating viral spike immunopeptides presented via MHC-class II (MHC-II) molecules by dendritic cells pulsed with SARS-CoV-2 spike protein, but it remains unknown if endogenous immunopep ...[more]

PMID: 38022636

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