Studies on the "insoluble" glycoprotein complex from human colon. Identification of reduction-insensitive MUC2 oligomers and C-terminal cleavage.
The "insoluble" glycoprotein complex was isolated from human colonic tissue and mucin subunits were prepared following reduction. Antibodies raised against peptide sequences within MUC2 revealed that virtually all of this mucin occurs in the insoluble glycoprotein complex. In addition, reduction released a 120-kDa C-terminal MUC2 fragment, showing that proteolytic cleavage in this domain may occur and leave the fragment attached to the complex via disulfide bonds. The variable number tandem repeat region and the irregular repeat domain were isolated after trypsin digestion and shown to have molecular weights of 930,000 and 180,000, respectively, suggesting a molecular weight for the entire MUC2 monomer of approximately 1.5 million. Gel chromatography and agarose gel electrophoresis revealed several populations of MUC2 subunits, and analytical ultracentrifugation showed that these have molecular weights on the order of 2 million, 4 million, and 5 million, corresponding to monomers, dimers, and trimers, respectively. Agarose gel electrophoresis of subunits from individuals expressing both a "long" and a "short" MUC2 allele revealed a larger number of populations, consistent with the presence of short and long monomers and oligomers arising from permutations of the two types of monomers. In addition to disulfide bonds, MUC2 monomers are apparently joined by a "novel," reduction-insensitive bond.
Herrmann
A
A
Mucosal Biology Group, Department of Cell and Molecular Biology, Section for Molecular Pathogenesis, Lund University, P. O. Box 94, 221 00 Lund, Sweden.
Davies
J R
JR
Lindell
G
G
Mårtensson
S
S
Packer
N H
NH
Swallow
D M
DM
Carlstedt
I
I
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000906:Antibodies / Q000378:metabolism; D003106:Colon / Q000737:chemistry*; D004220:Disulfides / Q000737:chemistry; D006020:Glycopeptides / Q000737:chemistry; D006023:Glycoproteins / Q000737:chemistry / Q000276:immunology; D006801:Humans; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009005:Monosaccharides / Q000032:analysis; D055262:Mucin-2; D009077:Mucins / Q000737:chemistry* / Q000276:immunology; D012995:Solubility; D014357:Trypsin / Q000378:metabolism
D000596:Amino Acids; D000906:Antibodies; D004220:Disulfides; D006020:Glycopeptides; D006023:Glycoproteins; C524625:MUC2 protein, human; D009005:Monosaccharides; D055262:Mucin-2; D009077:Mucins; D014357:Trypsin