sodium-dodecyl-sulfate and Teratoma

The tumor-associated cell surface antigens (TAA) gp 71 and AgI cocap H-2, but are not associated with H-2 following solubilization of membrane with detergent. We have attempted to solubilize intact TAA/H-2 complexes from surface-labeled L cells using decreasing concentrations of the nonionic detergent Nonidet-P 40 (NP 40). Associations between gp 71 and H-2 and between AgI and H-2 in solution were detected using sequential immunoprecipitation. gp 71/H-2 and AgI/H-2 complexes were solubilized at the lowest detergent concentration used, 0.01% NP 40. No coprecipitation of these antigen pairs was seen at any higher NP 40 concentration tested. Sodium dodecyl sulfate polyacrylamide gel electrophoresis revealed lthat precipitation of antigens solulbilized in 0.01% NP 40 leads to coprecipitation of a small number of other protein peaks and does not appear to involve wholesale inclusion of labeled membrane proteins. These results, together with previous observations of cocapping between TAA and H-2 antigens, suggest that the molecules are associated with one another in the membrane. Detergent solubilization of membranes with all but the most gentle conditions leads to disruption of these complexes.

Topics: Animals; Antigens, Neoplasm; Antigens, Surface; Cell Line; Electrophoresis, Polyacrylamide Gel; H-2 Antigens; Immunologic Capping; L Cells; Mice; Neoplasms, Experimental; Octoxynol; Polyethylene Glycols; Rabbits; Sodium Dodecyl Sulfate; Teratoma