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Function of the CysD domain of the gel-forming MUC2 mucin

The colonic human MUC2 mucin forms a polymeric gel by covalent disulfide bonds in its N- and C-termini. The middle part of MUC2 is largely composed of two highly O-glycosylated mucin domains that are interrupted by a CysD domain of unknown function. We studied its function as recombinant proteins fu...

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Autores principales: Ambort, Daniel, van der Post, Sjoerd, Johansson, Malin E. V., MacKenzie, Jenny, Thomsson, Elisabeth, Krengel, Ute, Hansson, Gunnar C.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Portland Press Ltd. 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3195396/
https://www.ncbi.nlm.nih.gov/pubmed/21338337
http://dx.doi.org/10.1042/BJ20102066
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author Ambort, Daniel
van der Post, Sjoerd
Johansson, Malin E. V.
MacKenzie, Jenny
Thomsson, Elisabeth
Krengel, Ute
Hansson, Gunnar C.
author_facet Ambort, Daniel
van der Post, Sjoerd
Johansson, Malin E. V.
MacKenzie, Jenny
Thomsson, Elisabeth
Krengel, Ute
Hansson, Gunnar C.
author_sort Ambort, Daniel
collection PubMed
description The colonic human MUC2 mucin forms a polymeric gel by covalent disulfide bonds in its N- and C-termini. The middle part of MUC2 is largely composed of two highly O-glycosylated mucin domains that are interrupted by a CysD domain of unknown function. We studied its function as recombinant proteins fused to a removable immunoglobulin Fc domain. Analysis of affinity-purified fusion proteins by native gel electrophoresis and gel filtration showed that they formed oligomeric complexes. Analysis of the individual isolated CysD parts showed that they formed dimers both when flanked by two MUC2 tandem repeats and without these. Cleavages of the two non-reduced CysD fusion proteins and analysis by MS revealed the localization of all five CysD disulfide bonds and that the predicted C-mannosylated site was not glycosylated. All disulfide bonds were within individual peptides showing that the domain was stabilized by intramolecular disulfide bonds and that CysD dimers were of non-covalent nature. These observations suggest that CysD domains act as non-covalent cross-links in the MUC2 gel, thereby determining the pore sizes of the mucus.
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spelling pubmed-31953962011-12-23 Function of the CysD domain of the gel-forming MUC2 mucin Ambort, Daniel van der Post, Sjoerd Johansson, Malin E. V. MacKenzie, Jenny Thomsson, Elisabeth Krengel, Ute Hansson, Gunnar C. Biochem J Research Article The colonic human MUC2 mucin forms a polymeric gel by covalent disulfide bonds in its N- and C-termini. The middle part of MUC2 is largely composed of two highly O-glycosylated mucin domains that are interrupted by a CysD domain of unknown function. We studied its function as recombinant proteins fused to a removable immunoglobulin Fc domain. Analysis of affinity-purified fusion proteins by native gel electrophoresis and gel filtration showed that they formed oligomeric complexes. Analysis of the individual isolated CysD parts showed that they formed dimers both when flanked by two MUC2 tandem repeats and without these. Cleavages of the two non-reduced CysD fusion proteins and analysis by MS revealed the localization of all five CysD disulfide bonds and that the predicted C-mannosylated site was not glycosylated. All disulfide bonds were within individual peptides showing that the domain was stabilized by intramolecular disulfide bonds and that CysD dimers were of non-covalent nature. These observations suggest that CysD domains act as non-covalent cross-links in the MUC2 gel, thereby determining the pore sizes of the mucus. Portland Press Ltd. 2011-04-27 2011-05-15 /pmc/articles/PMC3195396/ /pubmed/21338337 http://dx.doi.org/10.1042/BJ20102066 Text en © 2011 The Author(s) The author(s) has paid for this article to be freely available under the terms of the Creative Commons Attribution Non-Commercial Licence (http://creativecommons.org/licenses/by-nc/2.5/) which permits unrestricted non-commercial use, distribution and reproduction in any medium, provided the original work is properly cited. http://creativecommons.org/licenses/by-nc/2.5/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Ambort, Daniel
van der Post, Sjoerd
Johansson, Malin E. V.
MacKenzie, Jenny
Thomsson, Elisabeth
Krengel, Ute
Hansson, Gunnar C.
Function of the CysD domain of the gel-forming MUC2 mucin
title Function of the CysD domain of the gel-forming MUC2 mucin
title_full Function of the CysD domain of the gel-forming MUC2 mucin
title_fullStr Function of the CysD domain of the gel-forming MUC2 mucin
title_full_unstemmed Function of the CysD domain of the gel-forming MUC2 mucin
title_short Function of the CysD domain of the gel-forming MUC2 mucin
title_sort function of the cysd domain of the gel-forming muc2 mucin
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3195396/
https://www.ncbi.nlm.nih.gov/pubmed/21338337
http://dx.doi.org/10.1042/BJ20102066
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