Evaluation of MUC6 mucin tandem repeats

Parry, Simon and Sutton-Smith, Mark and Heal, Paul and Leir, Shih-Hsing and Palmai-Pallag, Timea and Morris, Howard R. and Hollingsworth, Michael A. and Dell, Anne and Harris, Ann (2005) Evaluation of MUC6 mucin tandem repeats. BBA - Biochimica et Biophysica Acta - General Subjects, 1722 (1). pp. 77-83. ISSN 0006-3002

Full content URL: http://dx.doi.org/10.1016/j.bbagen.2004.11.010

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Evaluation of MUC6 mucin tandem repeats

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Abstract

The MUC6 mucin was originally isolated from stomach mucus and is one of the major secreted mucins of the digestive tract. A full-length cDNA has not been isolated for this large molecule (greater than 15 kb) and it remains poorly studied. To circumvent the lack of reagents for investigating MUC6, we isolated a cDNA clone from a human fetal pancreatic duct cDNA library that encodes 282 amino acids of the MUC6 tandem repeat. A blast search with the sequence of this cDNA clone showed 90% homology with the original MUC6 (L07517) derived from a human stomach cDNA library and 95% homology both with AK096772, a MUC6-related protein isolated from a human prostate cDNA library and the human genome project clone AC083984. The MUC6 partial cDNA clone isolated from fetal pancreas was inserted into an epitope-tagged MUC1 mucin molecule in place of the native tandem repeat. This chimeric mucin was expressed in human pancreatic (Panc1) and colon (Caco2) carcinoma cell lines and purified for analysis of O-glycosylation by fast atom bombardment mass spectrometry (FAB-MS). The FAB-MS spectra showed O-glycans that had been detected previously on chimeric mucins carrying different tandem repeats, though the spectra for MUC1F/6TR mucins expressed in the Panc1 and Caco2 cells were very different. There was a paucity of O-glycosylation in Panc1 cells in comparison to Caco2 cells where many more structures were evident, and the most abundant glycans in Panc1 cells were sialylated.

Keywords:MUC6 mucin, Pancreas, Panc1 cell, Caco2 cell, Fast atom bombardment mass spectrometry
Subjects:C Biological Sciences > C130 Cell Biology
C Biological Sciences > C700 Molecular Biology, Biophysics and Biochemistry
Divisions:College of Science > School of Life Sciences
ID Code:16003
Deposited On:14 Nov 2014 09:14

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