The N terminus of the MUC2 mucin forms trimers that are held together within a trypsin-resistant core fragment
(2002) In Journal of Biological Chemistry 277(49). p.47248-47256- Abstract
- The N terminus of the human MUC2 mucin (amino acids 1-1397) has been expressed as a recombinant tagged protein in Chinese hamster ovary cells. The intracellular form was found to be an endoglycosidase H-sensitive monomer, whereas the secreted form was an oligomer that gave monomers upon disulfide bond reduction. The secreted MUC2 N terminus contained a trypsin-resistant core fragment. Edman sequencing and mass spectrometry of the peptides obtained localized this core fragment to the C-terminal end of the recombinant protein. This core retained its oligomeric nature with an apparent mass of similar to240 kDa. Upon reduction, peptides of similar to85 kDa were found, suggesting that the N terminus forms trimers. This interpretation was also... (More)
- The N terminus of the human MUC2 mucin (amino acids 1-1397) has been expressed as a recombinant tagged protein in Chinese hamster ovary cells. The intracellular form was found to be an endoglycosidase H-sensitive monomer, whereas the secreted form was an oligomer that gave monomers upon disulfide bond reduction. The secreted MUC2 N terminus contained a trypsin-resistant core fragment. Edman sequencing and mass spectrometry of the peptides obtained localized this core fragment to the C-terminal end of the recombinant protein. This core retained its oligomeric nature with an apparent mass of similar to240 kDa. Upon reduction, peptides of similar to85 kDa were found, suggesting that the N terminus forms trimers. This interpretation was also supported by gel electrophoresis and gel filtration of the intact MUC2 N terminus. Electron microscopy revealed three globular domains each linked via an extended and flexible region to a central part in a trefoil-like manner. Immunostaining with gold-labeled antibodies localized the N-terminal end to the three globular structures, and the antibodies directed against the Myc and green fluorescent protein tags attached at the C terminus localized these to the stalk side of the central trefoil. The N terminus of the MUC2 mucin is thus assembled into trimers that contain proteolytically stable parts, suggesting that MUC2 can only be partly degraded by intestinal proteases and thus is able to maintain a mucin network protecting the intestine. (Less)
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https://lup.lub.lu.se/record/321652
- author
- Godl, K ; Johansson, MEV ; Lidell, ME ; Mörgelin, Matthias LU ; Karlsson, H ; Olson, FJ ; Gum, JR ; Kim, YS and Hansson, GC
- organization
- publishing date
- 2002
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Journal of Biological Chemistry
- volume
- 277
- issue
- 49
- pages
- 47248 - 47256
- publisher
- American Society for Biochemistry and Molecular Biology
- external identifiers
-
- wos:000179663700053
- scopus:0037033038
- pmid:12374796
- ISSN
- 1083-351X
- DOI
- 10.1074/jbc.M208483200
- language
- English
- LU publication?
- yes
- id
- e07d0d94-76cf-4ed5-8dcd-aa321504593a (old id 321652)
- date added to LUP
- 2016-04-01 12:01:08
- date last changed
- 2022-04-05 08:23:17
@article{e07d0d94-76cf-4ed5-8dcd-aa321504593a,
abstract = {{The N terminus of the human MUC2 mucin (amino acids 1-1397) has been expressed as a recombinant tagged protein in Chinese hamster ovary cells. The intracellular form was found to be an endoglycosidase H-sensitive monomer, whereas the secreted form was an oligomer that gave monomers upon disulfide bond reduction. The secreted MUC2 N terminus contained a trypsin-resistant core fragment. Edman sequencing and mass spectrometry of the peptides obtained localized this core fragment to the C-terminal end of the recombinant protein. This core retained its oligomeric nature with an apparent mass of similar to240 kDa. Upon reduction, peptides of similar to85 kDa were found, suggesting that the N terminus forms trimers. This interpretation was also supported by gel electrophoresis and gel filtration of the intact MUC2 N terminus. Electron microscopy revealed three globular domains each linked via an extended and flexible region to a central part in a trefoil-like manner. Immunostaining with gold-labeled antibodies localized the N-terminal end to the three globular structures, and the antibodies directed against the Myc and green fluorescent protein tags attached at the C terminus localized these to the stalk side of the central trefoil. The N terminus of the MUC2 mucin is thus assembled into trimers that contain proteolytically stable parts, suggesting that MUC2 can only be partly degraded by intestinal proteases and thus is able to maintain a mucin network protecting the intestine.}},
author = {{Godl, K and Johansson, MEV and Lidell, ME and Mörgelin, Matthias and Karlsson, H and Olson, FJ and Gum, JR and Kim, YS and Hansson, GC}},
issn = {{1083-351X}},
language = {{eng}},
number = {{49}},
pages = {{47248--47256}},
publisher = {{American Society for Biochemistry and Molecular Biology}},
series = {{Journal of Biological Chemistry}},
title = {{The N terminus of the MUC2 mucin forms trimers that are held together within a trypsin-resistant core fragment}},
url = {{http://dx.doi.org/10.1074/jbc.M208483200}},
doi = {{10.1074/jbc.M208483200}},
volume = {{277}},
year = {{2002}},
}