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Accessibility of plasma membrane antigens
Authors:Sohan L. Gupta  Gideon Goldstein  Edward A. Boyse
Affiliation:(1) Memorial Sloan-Kettering Cancer Center, 10021 New York, New York
Abstract:Serological techniques applied to intact cells register only those antigens of the plasma membrane that are exposed at the cell surface and are therefore accessible to antibody. Solubilization of the plasma membrane by detergent, used in the conventional surface-iodination immunoprecipitation technique, renders other plasma membrane antigens accessible. We have shown this by using a modified version of the technique in which lysis with detergent is postponed until after the cells have been reacted with antibody. Comparison of the conventional and modified methods confirms that the plasma membrane glycoprotein gp70 has antigen that is not exposed on the intact cells as well as accessible antigen, for example, GIX. The modified surface-iodination immunoprecipitation method is useful for distinguishing cell-surface antigens from plasma membrane antigens that normally are not accessible. This is exemplified by the fact that standard anti-TL and anti-X.1 sera identify gp70 antigen in the plasma membrane that is registered by the conventional, but not by the modified method.Abbreviations used in this paper are agr anti - BALB BALB/c - gp70 MuLV envelope glycoprotein of molecular weight about 70,000 daltons, sometimes referred to as gp69/71 - gs group-specific - 125I-imm-pptn surface labeling of viable cells with125I followed by immunoprecipitation analysis - Ig immunoglobulin - MuLV murine leukemia virus - NMS normal mouse serum - PAGE polyacrylamide gel electrophoresis - PBS Dulbecco's phosphate-buffered saline, Ca++- and Mg++-free - SDS sodium dodecyl sulfate - TL thymus leukemia antigen
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