Commandeur U, Koenig R, Lesemann D E, Torrance L, Burgermeister W, Liu Y, Schots A, Alric M, Grassi G
Biologische Bundesanstalt für Land- und Forstwirtschaft, Institut für Biochemie und Pflanzenvirologie, Braunschweig, Germany.
J Gen Virol. 1992 Mar;73 ( Pt 3):695-700. doi: 10.1099/0022-1317-73-3-695.
The location of five SDS-stable epitopes on the coat protein (CP) of beet necrotic yellow vein virus was determined by reacting Escherichia coli-expressed free CP, as well as fusion proteins (FP) containing fragments of the CP, with polyclonal and monoclonal antibodies on Western blots. Epitope 1, which has previously been found to be exposed on only one extremity of the virus particle, was located in the region between amino acids (aa) 1 and 7, i.e. on the N terminus of the CP. It was blocked when the N terminus of the CP was linked to a portion of the beta-galactosidase sequence in an FP. Epitope 3, which has previously been found to be exposed on the opposite extremity of the particle, was located in the region between aa 37 and 59. Epitope 4, which is exposed along the entire length of the particle, occurs on the C terminus of CP (aa 183 to 188). Two previously unknown epitopes were identified in the regions between aa 115 and 125 and 125 and 140, respectively. The former was located on the same extremity of the particle as epitope 3, the latter became accessible only after denaturation of the particle. Nothing is known about the probably non-adjacent aa sequences that participate in the formation of the two SDS-labile epitopes (epitopes 2 and 5) which are found on one extremity and along the entire length of the particle, respectively.
通过在蛋白质免疫印迹中使大肠杆菌表达的游离外壳蛋白（CP）以及含有CP片段的融合蛋白（FP）与多克隆抗体和单克隆抗体反应，确定了甜菜坏死黄脉病毒外壳蛋白上五个对十二烷基硫酸钠（SDS）稳定的表位的位置。表位1先前已发现仅暴露于病毒粒子的一个末端，位于氨基酸（aa）1至7之间的区域，即在CP的N末端。当CP的N末端与FP中的一部分β-半乳糖苷酶序列连接时，它被阻断。表位3先前已发现暴露于粒子的相对末端，位于aa 37至59之间的区域。表位4沿粒子的整个长度暴露，位于CP的C末端（aa 183至188）。在aa 115至125和125至140之间的区域分别鉴定出两个先前未知的表位。前者与表位3位于粒子的同一末端，后者仅在粒子变性后才变得可及。关于可能不相邻的氨基酸序列一无所知，这些序列分别参与了在粒子的一个末端和整个长度上发现的两个对SDS不稳定的表位（表位2和5）的形成。
