Enomoto T, Lichy J H, Ikeda J E, Hurwitz J
Proc Natl Acad Sci U S A. 1981 Nov;78(11):6779-83. doi: 10.1073/pnas.78.11.6779.
The 80,000-dalton form of the adenovirus (Ad) terminal protein (pTP) has been purified from Ad-infected HeLa cells. pTP was assayed by its ability to form a covalent complex with dCMP. The protein copurified with an activity that is essential for in vitro Ad DNA replication (Ad protein activity) as well as with a DNA polymerase activity that was distinguished from those of HeLa cell DNA polymerases alpha, beta, and gamma. The Ad protein-associated DNA polymerase activity was detected with activated DNA but not with poly(rA).oligo(dT) as template and was insensitive to aphidicolin and sensitive to N-ethylmaleimide. The Ad protein, DNA polymerase, and pTP-dCMP complex-forming activities sedimented in a glycerol gradient as a single peak with an apparent molecular size of 180,000 daltons. NaDodSO4/polyacrylamide gel analysis of the glycerol gradient fraction showed major bands of 80,000 and 140,000 daltons. The 80,000-dalton band was identified as pTP by comparison of its tryptic peptide map with that of the 55,000-dalton form of the terminal protein, which was purified from Ad virions.
已从腺病毒（Ad）感染的HeLa细胞中纯化出80,000道尔顿形式的腺病毒末端蛋白（pTP）。通过其与dCMP形成共价复合物的能力来检测pTP。该蛋白与体外Ad DNA复制所必需的一种活性（Ad蛋白活性）以及一种与HeLa细胞DNA聚合酶α、β和γ不同的DNA聚合酶活性共同纯化。Ad蛋白相关的DNA聚合酶活性在用活化DNA作为模板时可检测到，但以聚（rA）·寡聚（dT）作为模板时则检测不到，并且对阿非迪霉素不敏感，对N - 乙基马来酰亚胺敏感。Ad蛋白、DNA聚合酶和pTP - dCMP复合物形成活性在甘油梯度中以单一峰沉降，表观分子大小为180,000道尔顿。对甘油梯度级分进行的十二烷基硫酸钠/聚丙烯酰胺凝胶分析显示出80,000和140,000道尔顿的主要条带。通过将其胰蛋白酶肽图谱与从Ad病毒粒子中纯化的55,000道尔顿形式的末端蛋白的图谱进行比较，确定80,000道尔顿的条带为pTP。
