Koenig R, Commandeur U, Lesemann D E, Burgermeister W, Torrance L, Grassi G, Alric M, Kallerhoff J, Schots A
Biologische Bundesanstalt für Land- und Forstwirtschaft, Institute für Viruskrankheiten der Pflanzen und Biochemie, Braunschweig, F.R.G.
J Gen Virol. 1990 Oct;71 ( Pt 10):2229-32. doi: 10.1099/0022-1317-71-10-2229.
By means of monoclonal antibodies (MAbs), five (groups of) epitopes were identified on particles of beet necrotic yellow vein virus (BNYVV). Epitopes 1 and 2, which were located on the opposite extremities of virus particles, are discontinuous (SDS-labile) epitopes which were destroyed when the particles were treated with trypsin. Epitope 3 is a continuous (SDS-stable) epitope located at the same extremity as epitope 2. It was not destroyed when the particles were treated with trypsin and was present on an Escherichia coli-expressed fusion protein containing amino acids (aa) 1 to 103 of the BNYVV coat protein. The continuous epitope 4, which was located along the entire length of the particles, was found to be present on a fusion protein containing aa 104 to 188 of the BNYVV coat protein but not on trypsin-treated virus particles. In Western blots, these treated particles yielded two slightly smaller coat proteins which failed to react with MAbs specific for epitope 4 but did react with polyclonal antisera and MAbs specific for epitope 3. BNYVV coat protein has a trypsin cleavage site on the carboxyl side of arginine in position 182, so it is therefore suggested that epitope 4 is located on the exposed C terminus, which is composed of aa 183 to 188. Epitope 5 was also located along the entire length of the particles but in a more uneven distribution than epitope 4. This may be because it is a discontinuous epitope that is very sensitive to subtle changes in protein conformation.
通过单克隆抗体（MAbs），在甜菜坏死黄脉病毒（BNYVV）颗粒上鉴定出了五个（组）表位。表位1和表位2位于病毒颗粒的相对末端，是不连续（对SDS不稳定）的表位，在用胰蛋白酶处理颗粒时会被破坏。表位3是一个连续（对SDS稳定）的表位，与表位2位于同一末端。在用胰蛋白酶处理颗粒时它不会被破坏，并且存在于大肠杆菌表达的融合蛋白上，该融合蛋白包含BNYVV外壳蛋白的第1至103个氨基酸（aa）。连续的表位4沿着颗粒的全长分布，发现它存在于包含BNYVV外壳蛋白第104至188个氨基酸的融合蛋白上，但不存在于经胰蛋白酶处理的病毒颗粒上。在蛋白质免疫印迹中，这些经处理的颗粒产生了两种略小的外壳蛋白，它们不能与针对表位4的单克隆抗体发生反应，但能与针对表位3的多克隆抗血清和单克隆抗体发生反应。BNYVV外壳蛋白在第182位精氨酸的羧基侧有一个胰蛋白酶切割位点，因此推测表位4位于暴露的C末端，该末端由第183至188个氨基酸组成。表位5也沿着颗粒的全长分布，但分布比表位4更不均匀。这可能是因为它是一个不连续的表位，对蛋白质构象的细微变化非常敏感。
