Glycobiology Advance Access published online on June 25, 2007
Glycobiology, doi:10.1093/glycob/cwm068
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Mucin biosynthesis: Molecular cloning and expression of mouse mucus-type core 2 ß1,6 N-acetylglucosaminyltransferase*
1 the Department of Pathology, Nagoya University Graduate School of Medicine, Nagoya, Japan
2 the Department of Biochemistry and Molecular Biology, College of Medicine, University of Nebraska Medical Center
3 Eppley Cancer Center for Research in Cancer and Allied Diseases, University of Nebraska Medical Center, Omaha, NE 68198-5870
Address correspondence to: Dr. Pi-Wan Cheng, the Department of Biochemistry and Molecular Biology, College of Medicine, University of Nebraska Medical Center, 985870 Nebraska Medical Center, Omaha, NE 68198-5870, Tel. 402 559-5776; Fax, 402 559-6650; E-mail: pcheng{at}unmc.edu
Received on April 27, 2007; accepted on June 17, 2007
Secreted mucins protect the underlying epithelium by serving as the major determinant of the rheological property of mucus secretion and the receptors for pathogens. These functions can be affected by the three branch structures, including core 2, core 4, and blood group I, which are synthesized by the mucus-type core 2 ß1,6 N-acetylglucosaminyltransferase (C2GnT-M). Decreased activity of this enzyme and expression of this gene have been found in colorectal cancer, which supports the important role of this enzyme in the protective functions of secreted mucins. We cloned full-length mouse (m) C2GnT-M cDNAs and showed that the deduced amino acid sequence was homologous to those of other C2GnT-Ms. The recombinant protein generated by mC2GnT-M cDNA exhibited core 2, core 4, and blood group I enzyme activities with a ratio of 1.00:0.46:1.05. We identified two different size transcripts by rapid amplification of cDNA ends and RT-PCR. Derived from the 6.6 kb mC2GnT-M gene composed of three exons and two introns, these two transcripts were intronless and differed by the length of the 3'untranslated region. In addition, exon 2 was found to be heterogeneous in size. This gene was highly expressed in the gastrointestinal tract, including colon, stomach, and small intestine. Antibodies generated against mC2GnT-M identified this enzyme in the goblet cells and other mucus cells/glands. This report provides the basis for further characterization of the regulation of mC2GnT-M gene expression and the biological functions of this gene.
Key words: C2GnT-M gene / C2GnT-M gene expression / mucin biosynthesis / mucus cell IHC staining with C2GnT-M antibody / mucin glycan branch enzyme
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