Glycoproteins in human parotid saliva assessed by lectin probes after resolution by sodium dodecyl sulphate-polyacrylamide gel electrophoresis.
Human parotid salivary glycoproteins separated by gradient sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and electroblotted onto nitrocellulose have been investigated using a battery of biotinylated lectin probes of characterized sugar specificity. Lectin binding, detected on blots using avidin-biotin complex (ABC) and a chemiluminescence generating substrate, was recorded on photographic film and compared with the original fluorescein isothiocyanate (FITC) stained blots or with Coomassie Brilliant Blue R-250-stained gels run in parallel. A number of glycoprotein bands which were undetected by protein stains or the periodic acid Schiff reaction were revealed by lectins. Binding by lectins from Concanavalia ensiformis, Lens culinaris, Limax flavus, Phaseolus vulgaris, Ricinus communis, Triticum vulgaris, Lotus tetragonobulus and Ulex europaeus indicated that sialylated and fucosylated triantennary and bisected, N-linked complex sugar chains were present on many glycoproteins in addition to the major glycosylated proline-rich glycoprotein (GI). Binding with lectins from Arachis hypogaea and Dolichos biflorus indicated that the O-linked sugar chains were confined to the alpha-heavy chain of Ig A. Comparison of lectin binding in samples from five healthy individuals revealed differences in a number of glycoproteins in addition to the previously characterized G1 and CON 1/CON 2 polymorphisms and demonstrated that the H blood group antigen was expressed mainly on G1 in parotid saliva. This study will be used as a basis upon which to study salivary glycoproteins in diseases affecting parotid glands.
Secretory and Soft Tissue Research Unit, Department of Oral Pathology, King's College School of Medicine and Dentistry, London, UK.
G H
GH
Carpenter
G B
GB
Proctor
C L
CL
Pankhurst
R W
RW
Linden
D K
DK
Shori
X S
XS
Zhang
D000328:Adult; D002240:Carbohydrate Sequence; D004591:Electrophoresis, Polyacrylamide Gel; D005069:Evaluation Studies as Topic; D006023:Glycoproteins / Q000235:genetics*; D006801:Humans; D037102:Lectins; D015335:Molecular Probes; D008969:Molecular Sequence Data; D010306:Parotid Gland / Q000737:chemistry*; D010503:Periodic Acid-Schiff Reaction; D011110:Polymorphism, Genetic; D011485:Protein Binding; D012471:Salivary Proteins and Peptides / Q000235:genetics*
D006023:Glycoproteins; D037102:Lectins; D015335:Molecular Probes; D012471:Salivary Proteins and Peptides