Evidence for shared epitopes within the 'naked' protein domains of human mucus glycoproteins. A study performed by using polyclonal antibodies and electron microscopy.
Polyclonal antibodies were raised in rabbits towards reduced subunits of human cervical mucus glycoproteins. The reduced subunits almost completely inhibited the antiserum, whereas the intact mucins and the heavily glycosylated fragments obtained after digestion of reduced subunits with trypsin (T-domains) caused only partial inhibition. Periodate oxidation of intact mucins, reduced subunits and T-domains caused no effect on the antibody response, and fragments obtained by more extensive proteolysis of the reduced subunits (P-domains) showed no inhibitory activity. By using electron microscopy, antibodies from T-domain-adsorbed antisera were revealed as bound to cervical mucin reduced subunits, either directly or with colloidal gold-Protein A. Binding sites (100-150 nm apart) were observed at the ends and at internal positions of the reduced subunits. We conclude that the antibodies do not recognize carbohydrate structures but are directed to two kinds of protein epitopes, one shared by whole mucins, reduced subunits and T-domains, and the other specific to the reduced subunit fragment. The latter epitopes are 'cryptic' and are probably shielded within folded protein domains stabilized by disulphide bonds. Human bronchial, cervical, gastric and salivary mucus glycoproteins share some of these cryptic epitopes.
Sheehan
J K
JK
Department of Biochemistry and Molecular Biology, University of Manchester, U.K.
Boot-Handford
R P
RP
Chantler
E
E
Carlstedt
I
I
Thornton
D J
DJ
D000906:Antibodies; D002584:Cervix Uteri / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000032:analysis*; D005260:Female; D005753:Gastric Mucosa / Q000378:metabolism; D006801:Humans; D008168:Lung / Q000378:metabolism; D008854:Microscopy, Electron; D009077:Mucins / Q000276:immunology* / Q000648:ultrastructure; D009093:Mucus / Q000737:chemistry*; D012463:Saliva / Q000378:metabolism
D000906:Antibodies; D000939:Epitopes; D009077:Mucins