Advanced

The N terminus of the MUC2 mucin forms trimers that are held together within a trypsin-resistant core fragment

Godl, K; Johansson, MEV; Lidell, ME; Mörgelin, Matthias LU ; Karlsson, H; Olson, FJ; Gum, JR; Kim, YS and Hansson, GC (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)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Journal of Biological Chemistry
volume
277
issue
49
pages
47248 - 47256
publisher
ASBMB
external identifiers
  • wos:000179663700053
  • scopus:0037033038
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
2007-08-20 16:40:42
date last changed
2017-10-01 03:45:38
@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    = {ASBMB},
  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},
  volume       = {277},
  year         = {2002},
}