title: Functional protective role for mucin glycosylated repetitive domains.
abstract: Mucins carry out a number of protective roles, some of which are more easily studied than others. One mucin function is believed to be the protection of the mucosal epithelium against acidic and proteolytic damage in the stomach and intestines. In the present work, a portion of stomach mucin tandem repeat sequence (Muc6) was joined to the catalytic domain of a reporter enzyme [human milk cholesterol esterase (CE)] to determine whether the former can protect the latter protein from damage. This Muc6 domain replaced a unique series of glycosylated C-terminal repeats normally present in CE. The chimeric protein (CE/Muc6) was expressed in two different cell lines and its properties compared to recombinant full-length CE and a truncated version of CE which contained only the catalytic domain (CE/trunc). Results showed that both CE and CE/Muc6 were resistant to denaturation by acid and to proteolysis by pepsin at low pH values or by pancreatic proteases compared to CE/trunc. Thus, a stomach Muc6 domain is sufficient to confer stability on the CE catalytic domain, demonstrating a protective effect by a glycosylated mucin sequence.
authors.affiliation: School of Biological Sciences, The University of Auckland, New Zealand. k.loomes@auckland.ac.nz
authors.forename: K M
authors.initials: KM
authors.lastname: Loomes
authors.forename: H E
authors.initials: HE
authors.lastname: Senior
authors.forename: P M
authors.initials: PM
authors.lastname: West
authors.forename: A M
authors.initials: AM
authors.lastname: Roberton
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D020134:Catalytic Domain; D002417:Cattle; D002850:Chromatography, Gel; D006224:Cricetinae; D010450:Endopeptidases / Q000378:metabolism; D005753:Gastric Mucosa / Q000378:metabolism; D006031:Glycosylation; D006863:Hydrogen-Ion Concentration; D037102:Lectins / Q000378:metabolism; D008969:Molecular Sequence Data; D055272:Mucin-6; D009077:Mucins / Q000737:chemistry* / Q000378:metabolism; D010434:Pepsin A / Q000378:metabolism; D011489:Protein Denaturation; D011499:Protein Processing, Post-Translational; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012091:Repetitive Sequences, Nucleic Acid
chemical_list: D037102:Lectins; C524734:MUC6 protein, human; D055272:Mucin-6; D009077:Mucins; D011993:Recombinant Fusion Proteins; D010450:Endopeptidases; D010434:Pepsin A