title: Lectin binding studies of parotid salivary glycoproteins in Sjögren's syndrome.
abstract: Human parotid salivas were collected from patients with secondary Sjögren's syndrome and controls without disease or with drug-induced xerostomia. Parotid glycoproteins were separated by gradient sodium dodecyl sulphate gel electrophoresis (SDS-PAGE), electroblotted onto nitrocellulose membrane and probed with biotinylated lectins of characterised sugar specificities. The binding patterns of lectins from Maclura pomifera (MPA) and Arachis hypogaea (PNA) indicated that many parotid glycoproteins have sialylated O-linked glycans and that sialylation is not affected by disease. Binding by lectins from Ricinus communis (RCA-1), Limax flavus (LFA), Lotus tetragonolobus (LTA) and Ulex europaeus (UEA-1) appeared unaltered in secondary Sjögren's syndrome, suggesting no obvious change in N-glycosylation of parotid glycoproteins. Variations in binding patterns of most lectins was attributable to subject-to-subject variations in recognised polymorphic proteins. Dolichos biflorus agglutinin (DBA) consistently showed increased binding to a 75 kDa (Mr) protein in salivas from patients with secondary Sjögren's syndrome. The binding protein was identified as lactoferrin but found not to contain N-acetylgalactosamine, the sugar to which DBA binds. Binding of DBA to lactoferrin was dependent upon its saturation with iron, modified SDS-PAGE under nonreducing conditions resolved iron-free and iron-saturated lactoferrins and demonstrated increased levels of the iron-saturated form in secondary Sjögren's syndrome. Lectin binding studies of purified lactoferrins from saliva, milk, and polymorphonuclear neutrophils suggested that raised levels of lactoferrin in saliva originate from salivary cells and not from inflammatory cells. These results suggest that DBA binding provides greater specificity as an indicator of salivary gland disease than measurement of lactoferrin levels alone.
authors.affiliation: Secretory & Soft Tissue Research Unit, Guy's, King's & St Thomas' School of Dentistry, London, UK. guy.carpenter@kcl.ac.uk
authors.forename: G H
authors.initials: GH
authors.lastname: Carpenter
authors.forename: C L
authors.initials: CL
authors.lastname: Pankhurst
authors.forename: G B
authors.initials: GB
authors.lastname: Proctor
mesh_terms: D019921:Biotinylation; D004591:Electrophoresis, Polyacrylamide Gel; D006023:Glycoproteins / Q000378:metabolism*; D006031:Glycosylation; D006801:Humans; D007501:Iron / Q000378:metabolism; D007781:Lactoferrin / Q000378:metabolism; D037102:Lectins / Q000378:metabolism*; D019158:N-Acetylneuraminic Acid / Q000378:metabolism; D010306:Parotid Gland / Q000378:metabolism*; D019887:Peanut Agglutinin / Q000378:metabolism; D037121:Plant Lectins; D012463:Saliva / Q000737:chemistry*; D012859:Sjogren's Syndrome / Q000378:metabolism*; D014987:Xerostomia / Q000139:chemically induced / Q000378:metabolism
chemical_list: D006023:Glycoproteins; D037102:Lectins; D019887:Peanut Agglutinin; D037121:Plant Lectins; C036554:dolichos biflorus agglutinin; C034551:maclurin; D007501:Iron; D007781:Lactoferrin; D019158:N-Acetylneuraminic Acid