Linné T, Jörnvall H, Philipson L
Eur J Biochem. 1977 Jun 15;76(2):481-90. doi: 10.1111/j.1432-1033.1977.tb11618.x.
The virus-coded 72000-Mr 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-Mr DNA-binding protein is phosphorylated and the phosphate is covalently linked predominantly to serine. Analysis of tryptic digests of the 32P-labeled 72000-Mr 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 [35S]methionine-labeled 72000-Mr and 45000-Mr polypeptides are indistinguishable. The amino acid compositions of the 72000-Mr and 45000-Mr polypeptides show closely related distributions. An antiserum produced against the purified 72000-Mr DNA-binding protein precipitates both the 72000-Mr and the 45000-Mr 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.
通过二乙氨基乙基纤维素色谱法、选择性沉淀法和凝胶过滤法，已将来自2型腺病毒感染细胞的病毒编码72000道尔顿DNA结合蛋白纯化至同质。72000道尔顿DNA结合蛋白被磷酸化，且磷酸主要以共价键形式连接到丝氨酸上。对32P标记的72000道尔顿蛋白的胰蛋白酶消化产物分析表明，磷酸残基仅存在于一个肽段中。DNA结合组分含有另一种分子量约为45000的非磷酸化蛋白。[35S]甲硫氨酸标记的72000道尔顿和45000道尔顿多肽的胰蛋白酶肽图无法区分。72000道尔顿和45000道尔顿多肽的氨基酸组成显示出密切相关的分布。针对纯化的72000道尔顿DNA结合蛋白产生的抗血清可从腺病毒感染细胞的提取物中沉淀出72000道尔顿和45000道尔顿的蛋白。免疫荧光研究显示，DNA结合蛋白积聚在感染细胞核的特征性结构中。
