title: Gel-forming and cell-associated mucins: preparation for structural and functional studies.
abstract: Secreted and transmembrane mucins are important components of innate defence at the body's mucosal surfaces. The secreted mucins are large, polymeric glycoproteins, which are largely responsible for the gel-like properties of mucus secretions. The cell-tethered mucins, however, are monomeric but are typically composed of two subunits, a larger extracellular subunit which is heavily glycosylated while the smaller more sparsely glycosylated subunit has a short extracellular region, a single-pass transmembrane domain, and a cytoplasmic tail. These two families of mucins represent high-molecular-weight glycoproteins containing serine and threonine-rich domains that are the attachment sites for large numbers of O-glycans. The high-M               (                 r               ) and high sugar content have been exploited for the separation of mucins from the majority of components in mucus secretions. In this chapter, we describe current and well-established methods (caesium chloride density-gradient centrifugation, gel-filtration and anion-exchange chromatography, and agarose gel electrophoresis) for the extraction and purification of gel-forming and cell-surface mucins which can subsequently be used for a variety of structural and functional studies.
authors.affiliation: Department of Oral Biology, Faculty of Odontology, Malmö University, Malmö, SE, Sweden. Julia.Davies@mah.se
authors.forename: Julia R
authors.initials: JR
authors.lastname: Davies
authors.forename: Claes
authors.initials: C
authors.lastname: Wickström
authors.forename: David J
authors.initials: DJ
authors.lastname: Thornton
mesh_terms: D002478:Cells, Cultured; D004847:Epithelial Cells / Q000737:chemistry*; D005782:Gels / Q000737:chemistry*; D006801:Humans; D009077:Mucins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D011487:Protein Conformation
chemical_list: D005782:Gels; D009077:Mucins