Jeng Y H, Wold W S, Green M
J Virol. 1978 Oct;28(1):314-23. doi: 10.1128/JVI.28.1.314-323.1978.
We have identified an adenovirus type 2 (Ad2)-induced early glycopolypeptide with an apparent molecular weight of 20,000 to 21,000 (20/21K), as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The 20/21K polypeptide could be labeled in vivo with [(3)H]glucosamine. [(35)S]methionine- and [(3)H]-glucosamine-labeled 20/21K polypeptides bound to concanavalin A-Sepharose columns and were eluted with 0.2 M methyl-alpha-d-mannoside. The pulse-labeled polypeptide appeared as a sharp band with an apparent molecular weight of 21K, but after a chase it converted to multiple bands with an average molecular weight of 20K. This variability in electrophoretic mobility is consistent with glycosylation or deglycosylation of the 20/21K polypeptide. Analysis of the pulse and pulse-chase-labeled forms by using partial proteolysis indicated that the polypeptides were highly related chemically, but not identical. Most of the 20/21K polypeptide is localized in the cytoplasm fraction of infected cells lysed by Nonidet P-40. The 20/21K polypeptide and a 44K polypeptide, labeled with [(35)S]methionine or [(3)H]glucosamine in Ad2-infected human cells, were precipitated by a rat antiserum against an Ad2-transformed rat cell line (T2C4), but not by antisera against three other Ad2-transformed rat cell lines, or by serum from nonimmune rats. The partial proteolysis patterns of the 20/21K and the 44K polypeptides were indistinguishable, indicating that the two polypeptides are highly related, and suggesting that the 44K polypeptide might be a dimer of the 20/21K polypeptide. The 20/21K polypeptide was also induced in Ad2-early infected monkey and hamster cells. These results imply that the 20/21K polypeptide is synthesized in Ad2-infected human, monkey, and hamster cells, and in one but not all Ad2-transformed rat cells. Thus, the 20/21K polypeptide is probably viral coded rather than cell coded and viral induced.
我们鉴定出一种腺病毒2型（Ad2）诱导的早期糖基化多肽，通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳估计其表观分子量为20,000至21,000（20/21K）。20/21K多肽可在体内用[³H]葡萄糖胺进行标记。用[³⁵S]甲硫氨酸和[³H]葡萄糖胺标记的20/21K多肽与伴刀豆球蛋白A-琼脂糖柱结合，并用0.2M甲基-α-D-甘露糖苷洗脱。脉冲标记的多肽呈现为一条表观分子量为21K的清晰条带，但追踪后转变为多条平均分子量为20K的条带。这种电泳迁移率变化与20/21K多肽的糖基化或去糖基化一致。通过部分蛋白酶解分析脉冲和脉冲追踪标记形式表明，这些多肽在化学上高度相关，但不完全相同。大多数20/21K多肽定位于被Nonidet P-40裂解的感染细胞的细胞质部分。在Ad2感染的人细胞中用[³⁵S]甲硫氨酸或[³H]葡萄糖胺标记的20/21K多肽和44K多肽，可被抗Ad2转化大鼠细胞系（T2C4）的大鼠抗血清沉淀，但不能被抗其他三种Ad2转化大鼠细胞系的抗血清沉淀，也不能被非免疫大鼠的血清沉淀。20/21K和44K多肽的部分蛋白酶解模式无法区分，表明这两种多肽高度相关，并提示44K多肽可能是20/21K多肽的二聚体。20/21K多肽也可在Ad2早期感染的猴和仓鼠细胞中诱导产生。这些结果表明，20/21K多肽在Ad2感染的人、猴和仓鼠细胞中合成，并且在一种但不是所有Ad2转化的大鼠细胞中合成。因此，20/21K多肽可能是病毒编码而非细胞编码且由病毒诱导产生的。
