Evaluation of MUC6 mucin tandem repeats

Simon Parry, Mark Sutton-Smith, Paul Heal, Shih Hsing Leir, Timea Palmai-Pallag, Howard R. Morris, Michael A. Hollingsworth, Anne Dell, Ann Harris*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

5 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)77-83
Number of pages7
JournalBiochimica et Biophysica Acta - General Subjects
Volume1722
Issue number1
DOIs
StatePublished - Feb 11 2005

Keywords

  • Caco2 cell
  • Fast atom bombardment mass spectrometry
  • MUC6 mucin
  • Panc1 cell
  • Pancreas

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology

Fingerprint Dive into the research topics of 'Evaluation of MUC6 mucin tandem repeats'. Together they form a unique fingerprint.

Cite this