Increased Understanding of the Biochemistry and Biosynthesis of MUC2 and Other Gel-Forming Mucins Through the Recombinant Expression of Their Protein Domains
Open accessAbstract
The gel-forming mucins are large and heavily O-glycosylated proteins which build up mucus gels. The recombinant production of full-length gel-forming mucins has not been possible to date. In order to study mucin biosynthesis and biochemistry, we and others have taken the alternative approach of constructing different recombinant proteins consisting of one or several domains of these large proteins and expressing them separately in different cell lines. Using this approach, we have determined that MUC2, the intestinal gel-forming mucin, dimerizes via its C-terminal cysteine-knot domain and also trimerizes via one of the N-terminal von Willebrand D domains. Both of these interactions are disulfide bond mediated. Via this assembly, a molecular network is built by which the mucus gel is formed. Here we discuss not only the functional understanding obtained from studies of the recombinant proteins, but also highlight the difficulties encountered when these proteins were produced recombinantly. We often found an accumulation of the proteins in the ER and consequently no secretion. This was especially apparent when the cysteine-rich domains of the N- and C-terminal parts of the mucins were expressed. Other proteins that we constructed were either not secreted or not expressed at all. Despite these problems, the knowledge of mucin biosynthesis and assembly has advanced considerably through the studies of these recombinant proteins.
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Integrity signals
Registry
Cites
Statistics
Claims the paper makes
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
For MUC5AC and MUC5B, higher molecular weight aggregates have been detected using SDS-PAGE under both reducing and non-reducing conditions.
“For MUC5AC and MUC5B, higher molecular weight aggregates have been detected using SDS-PAGE, under both reducing and non-reducing conditions [29].”
From High level in vivo mucin-type glycosylation in Escherichia coli (2018) · cites this paper as [29]
The CysD Domains section reports higher-molecular-mass aggregates in SDS gel electrophoresis of recombinant MUC5AC and MUC5B CysD domains. The claim omits that domain-specific qualification and could be read as referring to the full mucins. The passage also explicitly applies 'both reducing and non-reducing conditions' to the predominantly monomeric migration, rather than separately specifying the conditions under which aggregates appeared. (LLM, conf=0.86)
Experimental — accuracy not yet measured
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
The self-check needs the paper’s full text.
Cites
Structure and Function of the Polymeric Mucins in Airways Mucus
Thornton, D. J., Rousseau, K., & McGuckin, M. A. (2007) · Annual Review of Physiology · doi:10.1146/annurev.physiol.70.113006.100702 · Cited by 824 · see citing works
Composition and functional role of the mucus layers in the intestine
Johansson, M., Ambort, D., Pelaseyed, T., Schutte, A. H., Gustafsson, J. K., Ermund, A., Subramani, D. B., Holmén‐Larsson, J., Thomsson, K. A., Bergström, J. H., et al. (2011) · Cellular and Molecular Life Sciences · doi:10.1007/s00018-011-0822-3 · Cited by 559 · see citing works
Self-citationThe protein disulfide isomerase AGR2 is essential for production of intestinal mucus
Park, S.-W., Zhen, G., Verhaeghe, C., Nakagami, Y., Nguyenvu, L. T., Barczak, A. J., Killeen, N. P., & Erle, D. J. (2009) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0808722106 · Cited by 418 · see citing works
Gel-forming mucins appeared early in metazoan evolution
Lang, T., Hansson, G. C., & Samuelsson, T. (2007) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0705984104 · Cited by 310 · see citing works
Self-citationSewell, R., Bäckström, M., Dalziel, M., Gschmeissner, S., Karlsson, H., Noll, T., Gätgens, J., Clausen, H., Hansson, G. C., Burchell, J., et al. (2005) · Journal of Biological Chemistry · doi:10.1074/jbc.m511826200 · Cited by 258 · see citing works
Self-citationO-Linked Glycosylation in the Mammary Gland: Changes that Occur During Malignancy
Burchell, J., Mungul, A., & Taylor‐Papadimitriou, J. (2001) · Journal of Mammary Gland Biology and Neoplasia · doi:10.1023/a:1011331809881 · Cited by 221 · see citing works
Godl, K., Johansson, M., Lidell, M. E., Mörgelin, M., Karlsson, H., Olson, F. J., Gum, J. R., Kim, Y. S., & Hansson, G. C. (2002) · Journal of Biological Chemistry · doi:10.1074/jbc.m208483200 · Cited by 212 · see citing works
Self-citationBjörkman, Y. A., Thomsson, K. A., Larsson, J. M. H., Ekerhovd, E., & Hansson, G. C. (2007) · Molecular & Cellular Proteomics · doi:10.1074/mcp.m600439-mcp200 · Cited by 184 · see citing works
Self-citationMüller, S., & Hanisch, F. (2002) · Journal of Biological Chemistry · doi:10.1074/jbc.m202921200 · Cited by 136 · see citing works
C-Mannosylation of MUC5AC and MUC5B Cys subdomains
Pérez-Vilar, J. (2004) · Glycobiology · doi:10.1093/glycob/cwh041 · Cited by 110 · see citing works
Bäckström, M., Thomas, L. I. N. K., Olson, F. J., Karlsson, H., Graham, R. A., Picco, G., Burchell, J., Taylor‐Papadimitriou, J., Noll, T., & Hansson, G. C. (2003) · Biochemical Journal · doi:10.1042/bj20031130 · Cited by 105 · see citing works
Self-citationFunction of the CysD domain of the gel-forming MUC2 mucin
Ambort, D., van der Post, S., Johansson, M. E. V., MacKenzie, J., Thomsson, E., Krengel, U., & Hansson, G. C. (2011) · Biochemical Journal · doi:10.1042/bj20102066 · Cited by 104 · see citing works
Self-citationLidell, M. E., Johansson, M., & Hansson, G. C. (2003) · Journal of Biological Chemistry · doi:10.1074/jbc.m210069200 · Cited by 102 · see citing works
Self-citationTwo Cys residues essential for von Willebrand factor multimer assembly in the Golgi
Purvis, A. R., Gross, J., Dang, L. T., Huang, R., Kapadia, M., Townsend, R. R., & Sadler, J. E. (2007) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0705175104 · Cited by 88 · see citing works
Porcine Submaxillary Mucin Forms Disulfide-linked Multimers through Its Amino-terminal D-domains
Perez-Vilar, J., Eckhardt, A. E., DeLuca, A., & Hill, R. L. (1998) · Journal of Biological Chemistry · doi:10.1074/jbc.273.23.14442 · Cited by 79 · see citing works
Lidell, M. E., Johansson, M., Mörgelin, M., Asker, N., Gum, J. R., Kim, Y. S., & Hansson, G. C. (2003) · Biochemical Journal · doi:10.1042/bj20030003 · Cited by 76 · see citing works
Self-citationKrieg, J., Gläsner, W., Vicentini, A. M., Doucey, M., Löffler, A., Heß, D., & Hofsteenge, J. (1997) · Journal of Biological Chemistry · doi:10.1074/jbc.272.42.26687 · Cited by 69 · see citing works
Cleavage in the GDPH sequence of the C-terminal cysteine-rich part of the human MUC5AC mucin
Lidell, M. E., & Hansson, G. C. (2006) · Biochemical Journal · doi:10.1042/bj20060443 · Cited by 59 · see citing works
Self-citationMUC6 mucin expression inhibits tumor cell invasion
Leir, S., & Harris, A. (2011) · Experimental Cell Research · doi:10.1016/j.yexcr.2011.07.021 · Cited by 49 · see citing works
Perez-Vilar, J., & Hill, R. L. (1998) · Journal of Biological Chemistry · doi:10.1074/jbc.273.12.6982 · Cited by 44 · see citing works
Bell, S. L., Xu, G., & Forstner, J. (2001) · Biochemical Journal · doi:10.1042/0264-6021:3570203 · Cited by 39 · see citing works
Bell, S. L., Khatri, I. A., Xu, G., & Forstner, J. (1998) · European Journal of Biochemistry · doi:10.1046/j.1432-1327.1998.2530123.x · Cited by 39 · see citing works
Domino, S. E., Hurd, E. A., Thomsson, K. A., Karnak, D. M., Larsson, J. M. H., Thomsson, E., Bäckström, M., & Hansson, G. C. (2009) · Glycoconjugate Journal · doi:10.1007/s10719-009-9234-0 · Cited by 38 · see citing works
Self-citationSihlbom, C., Härd, I. V. D., Lidell, M. E., Noll, T., Hansson, G. C., & Bäckström, M. (2008) · Glycobiology · doi:10.1093/glycob/cwn144 · Cited by 38 · see citing works
Self-citationCloning and Characterization of Human MUC19 Gene
Zhu, L., Lee, P., Yu, D., Tao, S., & Chen, Y. (2010) · American Journal of Respiratory Cell and Molecular Biology · doi:10.1165/rcmb.2010-0312oc · Cited by 31 · see citing works
Premaratne, P., Welén, K., Damber, J., Hansson, G. C., & Bäckström, M. (2010) · Tumor Biology · doi:10.1007/s13277-010-0114-9 · Cited by 18 · see citing works
Self-citation
Flagged first, then by citations
Cited by
Sorted by citations
Mucin-type O-glycans and their roles in intestinal homeostasis
Bergstrom, K. S. B., & Xia, L. (2013) · Glycobiology · doi:10.1093/glycob/cwt045 · Cited by 360 · see citing works
High level in vivo mucin-type glycosylation in Escherichia coli
Mueller, P., Gauttam, R., Raab, N., Handrick, R., Wahl, C., Leptihn, S., Zorn, M., Kussmaul, M., Scheffold, M., Eikmanns, B., et al. (2018) · Microbial Cell Factories · doi:10.1186/s12934-018-1013-9 · Cited by 22 · see citing works
- Authors
- Bäckström, M., Ambort, D., Thomsson, E., Johansson, M. E. V., & Hansson, G. C.
- Affiliations
- University of Gothenburg (SE)
- Corresponding author
- Bäckström, Malin
- Venue
- Molecular Biotechnology
- Published
- 2013-01-29
- Volume
- 54
- Issue
- 2
- Pages
- 250-256
- ISSN
- 1073-6085 (ISSN-L)
- 1559-0305
- PMC
- PMC4868133
- OpenAlex
- W2061973814
- Cited by
- 53
- References
- 26
- Citations by year
- 2026: 2 · 2025: 5 · 2024: 4 · 2023: 4 · 2022: 3 · 2021: 4 · 2020: 3 · 2019: 2 · 2018: 9 · 2017: 1 · 2016: 2 · 2015: 7 · 2014: 5 · 2013: 2
- FWCI
- 2.39
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Licence
- www.springer.com/tdm (tdm)
- Locations
- Molecular Biotechnology (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- Funders
- National Institute of Allergy and Infectious Diseases (U01AI095473)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Glycosylation and Glycoproteins Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Proteoglycans and glycosaminoglycans research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Viral Infectious Diseases and Gene Expression in Insects
- Keywords
- MUC2, gel-forming mucins, mucin production, recombinant protein expression, protein dimerization, protein trimerization, disulfide bonds, O-glycosylation, endoplasmic reticulum retention, protein secretion, intestinal mucin
- MeSH terms
- Animals, Biochemistry, Cell Line, Chlorocebus aethiops, Cricetulus, Cysteine, Gels, Glycosylation, Humans, Recombinant Proteins, CHO Cells, Protein Structure, Tertiary, Dimerization, COS Cells, Cell Line, Tumor, Mucin-2
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
