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N-glycosylation of mannose receptor (CD206) regulates glycan binding by C-type lectin domains
The macrophage mannose receptor (MR, CD206) is a transmembrane endocytic lectin receptor, expressed in selected immune and endothelial cells, and is involved in immunity and maintaining homeostasis. Eight of the ten extracellular domains of the MR are C-type lectin domains (CTLDs) which mediate the...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society for Biochemistry and Molecular Biology
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9672410/ https://www.ncbi.nlm.nih.gov/pubmed/36244450 http://dx.doi.org/10.1016/j.jbc.2022.102591 |
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author | Stavenhagen, Kathrin Mehta, Akul Y. Laan, Lisa Gao, Chao Heimburg-Molinaro, Jamie van Die, Irma Cummings, Richard D. |
author_facet | Stavenhagen, Kathrin Mehta, Akul Y. Laan, Lisa Gao, Chao Heimburg-Molinaro, Jamie van Die, Irma Cummings, Richard D. |
author_sort | Stavenhagen, Kathrin |
collection | PubMed |
description | The macrophage mannose receptor (MR, CD206) is a transmembrane endocytic lectin receptor, expressed in selected immune and endothelial cells, and is involved in immunity and maintaining homeostasis. Eight of the ten extracellular domains of the MR are C-type lectin domains (CTLDs) which mediate the binding of mannose, fucose, and GlcNAc in a calcium-dependent manner. Previous studies indicated that self-glycosylation of MR regulates its glycan binding. To further explore this structure–function relationship, we studied herein a recombinant version of mouse MR CTLD4-7 fused to human Fc-portion of IgG (MR-Fc). The construct was expressed in different glycosylation-mutant cell lines to study the influence of differential glycosylation on receptor glycan-binding properties. We conducted site-specific N- and O-glycosylation analysis and glycosylation site characterization using mass spectrometry by which several novel O-glycosylation sites were identified in mouse MR and confirmed in human full-length MR. This information guided experiments evaluating the receptor functionality by glycan microarray analysis in combination with glycan-modifying enzymes. Treatment of active MR-Fc with combinations of exoglycosidases, including neuraminidase and galactosidases, resulted in the loss of trans-binding (binding of MR CTLDs to non-MR glycans), due to unmasking of terminal, nonreducing GlcNAc in N-glycans of the MR CTLDs. Regalactosylation of N-glycans rescues mannose binding by MR-Fc. Our results indicate that glycans within the MR CTLDs act as a regulatory switch by masking and unmasking self-ligands, including terminal, nonreducing GlcNAc in N-glycans, which could control MR activity in a tissue- and cell-specific manner or which potentially affect bacterial pathogenesis in an immunomodulatory fashion. |
format | Online Article Text |
id | pubmed-9672410 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | American Society for Biochemistry and Molecular Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-96724102022-11-21 N-glycosylation of mannose receptor (CD206) regulates glycan binding by C-type lectin domains Stavenhagen, Kathrin Mehta, Akul Y. Laan, Lisa Gao, Chao Heimburg-Molinaro, Jamie van Die, Irma Cummings, Richard D. J Biol Chem Research Article The macrophage mannose receptor (MR, CD206) is a transmembrane endocytic lectin receptor, expressed in selected immune and endothelial cells, and is involved in immunity and maintaining homeostasis. Eight of the ten extracellular domains of the MR are C-type lectin domains (CTLDs) which mediate the binding of mannose, fucose, and GlcNAc in a calcium-dependent manner. Previous studies indicated that self-glycosylation of MR regulates its glycan binding. To further explore this structure–function relationship, we studied herein a recombinant version of mouse MR CTLD4-7 fused to human Fc-portion of IgG (MR-Fc). The construct was expressed in different glycosylation-mutant cell lines to study the influence of differential glycosylation on receptor glycan-binding properties. We conducted site-specific N- and O-glycosylation analysis and glycosylation site characterization using mass spectrometry by which several novel O-glycosylation sites were identified in mouse MR and confirmed in human full-length MR. This information guided experiments evaluating the receptor functionality by glycan microarray analysis in combination with glycan-modifying enzymes. Treatment of active MR-Fc with combinations of exoglycosidases, including neuraminidase and galactosidases, resulted in the loss of trans-binding (binding of MR CTLDs to non-MR glycans), due to unmasking of terminal, nonreducing GlcNAc in N-glycans of the MR CTLDs. Regalactosylation of N-glycans rescues mannose binding by MR-Fc. Our results indicate that glycans within the MR CTLDs act as a regulatory switch by masking and unmasking self-ligands, including terminal, nonreducing GlcNAc in N-glycans, which could control MR activity in a tissue- and cell-specific manner or which potentially affect bacterial pathogenesis in an immunomodulatory fashion. American Society for Biochemistry and Molecular Biology 2022-10-13 /pmc/articles/PMC9672410/ /pubmed/36244450 http://dx.doi.org/10.1016/j.jbc.2022.102591 Text en © 2022 The Authors https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Research Article Stavenhagen, Kathrin Mehta, Akul Y. Laan, Lisa Gao, Chao Heimburg-Molinaro, Jamie van Die, Irma Cummings, Richard D. N-glycosylation of mannose receptor (CD206) regulates glycan binding by C-type lectin domains |
title | N-glycosylation of mannose receptor (CD206) regulates glycan binding by C-type lectin domains |
title_full | N-glycosylation of mannose receptor (CD206) regulates glycan binding by C-type lectin domains |
title_fullStr | N-glycosylation of mannose receptor (CD206) regulates glycan binding by C-type lectin domains |
title_full_unstemmed | N-glycosylation of mannose receptor (CD206) regulates glycan binding by C-type lectin domains |
title_short | N-glycosylation of mannose receptor (CD206) regulates glycan binding by C-type lectin domains |
title_sort | n-glycosylation of mannose receptor (cd206) regulates glycan binding by c-type lectin domains |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9672410/ https://www.ncbi.nlm.nih.gov/pubmed/36244450 http://dx.doi.org/10.1016/j.jbc.2022.102591 |
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