Karshin W L, Arcement L J, Naso R B, Arlinghaus R B
J Virol. 1977 Sep;23(3):787-98. doi: 10.1128/JVI.23.3.787-798.1977.
Rauscher murine leukemia virus glycoprotein gp69/71 and non-glycosylated p15(E) are synthesized by way of a 90,000-dalton precursor glycoprotein, termed Pr2a+b. Peptide mapping experiments showed that Pr2a+b contains all the tyrosine-containing tryptic peptides of gp69/71. Two additional tyrosine-containing tryptic peptides in Pr2a+b that are not detected in gp69/71 are found in p15(E). Thus, gp69/71 and p15(E) peptide sequences account for all the tyrosine tryptic peptides of Pr2a+b. The gene order of the two proteins was determined by pulse-labeling infected cells in the presence and absence of pactamycin at concentrations of the inhibitor that prevent initiation of translation, but not elongation. The gene order was found to be: (2)HN-gp69/71-p15(E)-COOH. A newly identified major viral protein, termed p12(E), migrates in sodium dodecyl sulfate-polyacrylamide gels in the "p12" region. It is related to p15(E) as determined by tryptic mapping experiments. p15(E) and p12(E) are not phosphorylated, and both can be separated from phosphoprotein p12 by guanidine hydrochloride-agarose chromatography. p12(E) and p15(E) elute in the void volume fraction, whereas phosphoprotein p12 elutes between p15 and p10. The two p12 proteins can also be separated from each other by two-dimensional gel electrophoresis involving isoelectric focusing in the first dimension and sodium dodecyl sulfate-gel electrophoresis in the second dimension.
劳舍尔鼠白血病病毒糖蛋白gp69/71和非糖基化的p15(E)是通过一种90,000道尔顿的前体糖蛋白Pr2a+b合成的。肽图谱实验表明,Pr2a+b包含gp69/71所有含酪氨酸的胰蛋白酶肽段。在p15(E)中发现了Pr2a+b中另外两个在gp69/71中未检测到的含酪氨酸的胰蛋白酶肽段。因此,gp69/71和p15(E)的肽序列构成了Pr2a+b的所有酪氨酸胰蛋白酶肽段。通过在存在和不存在放线菌酮的情况下脉冲标记感染细胞来确定这两种蛋白质的基因顺序,放线菌酮的浓度要能阻止翻译起始但不阻止延伸。发现基因顺序为:(2)HN-gp69/71-p15(E)-COOH。一种新鉴定的主要病毒蛋白,称为p12(E),在十二烷基硫酸钠-聚丙烯酰胺凝胶中迁移到“p12”区域。通过胰蛋白酶图谱实验确定它与p15(E)相关。p15(E)和p12(E)不被磷酸化,并且两者都可以通过盐酸胍-琼脂糖色谱与磷蛋白p12分离。p12(E)和p15(E)在空体积部分洗脱,而磷蛋白p12在p15和p10之间洗脱。这两种p12蛋白也可以通过二维凝胶电泳彼此分离,二维凝胶电泳包括在第一维进行等电聚焦,在第二维进行十二烷基硫酸钠-凝胶电泳。