Open AccessPurification and Characterization of the Phosphorylated DNA‐Binding Protein from Adenovirus‐Type‐2‐Infected Cells
Author(s) -
LINNÉ Tommy,
JÖRNVALL Hans,
PHILIPSON Lennart
Publication year - 1977
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1977.tb11618.x
Subject(s) - microbiology and biotechnology , dna , antiserum , biology , protein a/g , protein g , binding protein , biochemistry , antibody , recombinant dna , fusion protein , gene , immunology
The virus‐coded 72000‐ M r DNA‐binding protein from adenovirus‐type‐2‐infected cells has been purified to homogeneity by DEAE‐cellulose chromatography, selective precipitation and gel filtration. The 72000‐ M r DNA‐binding protein is phosphorylated and the phosphate is covalently linked predominantly to serine. Analysis of tryptic digests of the 32 P‐labeled 72000‐ M r protein showed that the phosphate residue(s) is present in only one peptide. The DNA‐binding fraction contains an additional non‐phosphorylated protein with an approximate molecular weight of 45000. Tryptic peptide maps of [ 35 S]methionine‐labeled 72000‐ M r and 45000‐ M r polypeptides are indistinguishable. The amino acid compositions of the 72000‐ M r and 45000‐ M r polypeptides show closely related distributions. An antiserum produced against the purified 72000‐ M r DNA‐binding protein precipitates both the 72000‐ M r and the 45000‐ M r protein from extracts of adenovirus‐infected cells. Immunofluorescence studies revealed DNA‐binding protein to be accumulated in characteristic structures in nuclei of the infected cells.