Human milk-fat globule membrane derived mucin is a disulfide-linked heteromer.
The human milk-fat globule membrane (HMFG) contains a number of antigens also expressed on breast tumors. The dominant antigens are a mucin of MW greater than 400,000 and a group of antigens of MW 67,000-70,000. The mucin was separated with monoclonal antibodies against HMFG and a MW 70,000 doublet was found to be associated with the mucin under non-reducing conditions, the latter identifiable by another set of monoclonal antibodies. Immunoprecipitation of the mucin and analysis of the precipitated material using Western blots and identification of transferred material with monoclonal antibodies demonstrated that the MW 70,000 protein and the mucin were co-precipitated and linked by reducible disulfide bonds. Treatment with detergents, protein-dissociating agents and glycosidases did not release the component from the mucin. Mucins in HMFG membrane and breast tumor cells may be associated therefore to disulfide-linked linker proteins similar to those described for intestinal and gastric mucins, that would co-purify with the mucin under non-reducing conditions.
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John Muir Cancer & Aging Research Institute, Walnut Creek, CA 94596.
Ceriani
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D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm; D015153:Blotting, Western; D019791:Guanidine; D006146:Guanidines; D006596:Hexosaminidases / Q000494:pharmacology; D006801:Humans; D007163:Immunosorbent Techniques; D008562:Membrane Glycoproteins / Q000032:analysis*; D008970:Molecular Weight; D018396:Mucin-1; D009077:Mucins / Q000032:analysis*; D009439:Neuraminidase / Q000494:pharmacology; D048809:alpha-N-Acetylgalactosaminidase
D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm; D006146:Guanidines; D008562:Membrane Glycoproteins; D018396:Mucin-1; D009077:Mucins; D006596:Hexosaminidases; D009439:Neuraminidase; C482722:NAGA protein, human; D048809:alpha-N-Acetylgalactosaminidase; D019791:Guanidine