Purification of a native membrane-associated adenovirus tumor antigen

H Persson, M G Katze, L Philipson
1982 Journal of Virology  
A 15,000-dalton protein was purified from HeLa cells infected with adenovirus type 2. Proteins solubilized from a membrane fraction of lytically infected cells was used as the starting material for purification. Subsequent purification steps involved lentil-lectin, phosphocellulose, hydroxyapatite, DEAE-cellulose, and aminohexyl-Sepharose chromatographies. A monospecific antiserum, raised against the purified protein, immunoprecipitated a 15,000-dalton protein encoded in early-region E1B
more » ... y-region E1B (ElB/15K protein) of the adenovirus type 2 DNA. Tryptic finger print analysis revealed that the purified protein was identical to the ElB/15K protein encoded in the transforming part of the viral genome. The antiserum immunoprecipitated the ElB/15K protein from a variety of viral transformed cell lines isolated from humans, rats, or hamsters. The ElB/15K protein was associated with the membrane fraction of both lytically and virustransformed cell lines and could only be released by detergent treatment. Furthermore, a 11,000-to 12,000-dalton protein that could be precipitated with the anti-ElB/15K serum was recovered from membranes treated with trypsin or proteinase K, suggesting that a major part of the ElB/15K protein is protected in membrane vesicles. Translation of early viral mRNA in a cell-free system, supplemented with rough microsomes, showed that this protein was associated with the membrane fraction also in vitro. on May 9, 2020 by guest
doi:10.1128/jvi.42.3.905-917.1982 fatcat:7ub3gsedzjhrpahcxpt42rxnse