Solubilization of complexes between tumor-associated and H-2 antigens from a tumor cell plasma membrane

European Journal of Immunology
P J Honeycutt, L R Gooding

Abstract

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.

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Citations

May 1, 1983·Human Immunology·R L Sparrow, I F McKenzie
Dec 1, 1985·Proceedings of the National Academy of Sciences of the United States of America·M FehlmannN Brossette
Dec 1, 1980·Immunology Today·M J Owen, M J Crumpton
Mar 15, 1983·International Journal of Cancer. Journal International Du Cancer·L Schaffar-DeshayesJ P Levy

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