Wychowski C, van der Werf S, Siffert O, Crainic R, Bruneau P, Girard M
EMBO J. 1983;2(11):2019-24. doi: 10.1002/j.1460-2075.1983.tb01694.x.
Using nuclease Bal31, deletions were generated within the poliovirus type 1 cDNA sequences, coding for capsid polypeptide VP1, within plasmid pCW119. The fusion proteins expressed in Escherichia coli by the deleted plasmids reacted with rabbit immune sera directed against poliovirus capsid polypeptide VP1 (alpha VP1 antibodies). They also reacted with a poliovirus type 1 neutralizing monoclonal antibody C3, but reactivity was lost when the deletion extended up to VP1 amino acids 90-104. Computer analysis of the protein revealed a high local density of hydrophilic amino acid residues in the region of VP1 amino acids 93-103. A peptide representing the sequence of this region was chemically synthesized. Once coupled to keyhole limpet hemocyanin, this peptide was specifically immunoprecipitated by C3 antibodies. The peptide also inhibited the neutralization of poliovirus type 1 by C3 antibodies. We thus conclude that the neutralization epitope recognized by C3 is located within the region of amino acids 93-104 of capsid polypeptide VP1.
利用核酸酶Bal31,在质粒pCW119内编码衣壳多肽VP1的脊髓灰质炎病毒1型cDNA序列中产生缺失。缺失质粒在大肠杆菌中表达的融合蛋白与针对脊髓灰质炎病毒衣壳多肽VP1的兔免疫血清(α VP1抗体)发生反应。它们也与脊髓灰质炎病毒1型中和单克隆抗体C3发生反应,但当缺失延伸至VP1氨基酸90 - 104时,反应性丧失。对该蛋白的计算机分析显示,在VP1氨基酸93 - 103区域存在高局部密度的亲水性氨基酸残基。合成了代表该区域序列的肽。该肽与钥孔戚血蓝蛋白偶联后,被C3抗体特异性免疫沉淀。该肽也抑制C3抗体对脊髓灰质炎病毒1型的中和作用。因此我们得出结论,C3识别的中和表位位于衣壳多肽VP1的氨基酸93 - 104区域内。