Definition of the primary structure of hepatitis B virus (HBV) pre-S hepatocyte binding domain using random peptide libraries
Definition of the primary structure of hepatitis B virus (HBV) pre-S hepatocyte binding domain using random peptide libraries
复制标题
DOI：
10.1006/viro.1997.8774
复制
发表时间：
1997-10-27
期刊：
影响因子：
3.7
通讯作者：
Howard, CR
中科院分区：
文献类型：
--
作者：
DMello, F;Partidos, CD;Howard, CR
前S特异性单克隆抗体MA 18/7已显示出抑制HBV与HepG 2细胞和肝膜的结合。因此，该抗体可用于鉴定参与肝细胞结合结构域的前S区的关键残基。使用重叠的7-mer肽代表HBV的前S区，由MA 18/7识别的表位显示含有来自前S1和前S2区的序列，从而表明肝细胞结合结构域是构象依赖性的。为了进一步研究前S蛋白上肝细胞结合区的一级结构，利用噬菌体展示的15肽库和8肽库分析了单克隆抗体MA 18/7的特异性。几个模拟表位被确定与噬菌体展示的肽库，其中大多数拥有一个中心的基序，至少有三个相同的残基内存在的天然前S1序列。当将这些模拟表位与前S2序列进行比较时，没有发现显著的共有序列。使用固相肽库鉴定的模拟表位也含有类似的基序。所有的噬菌体模拟表位和一个单一的模拟表位从固相肽库竞争与重组HBsAg颗粒含有前S1区结合MA 18/7。从噬菌体展示库中产生的针对四种模拟表位的小鼠抗血清与含有前S序列的HBsAg颗粒反应。这些数据表明，在前S区的保守的DPAF基序周围的前S分子的结构可能具有在结合HBV细胞受体的功能作用，并且在该区域的模拟表位中鉴定的中心基序可能为改进现有的甲型肝炎疫苗提供新的策略靶点，目前，这些疫苗大多缺乏前S特异性。(C)北京：科学出版社.
The pre-S-specific monoclonal antibody MA 18/7 has been shown to inhibit the binding of HBV to HepG2 cells and liver membranes. This antibody can thus be used to identify the critical residues of the pre-S region involved in the hepatocyte-binding domain. Using overlapping 7-mer peptides representing the pre-S region of HBV, the epitope recognized by MA 18/7 was shown to contain sequences from both the pre-S1 and pre-S2 regions, thus indicating that the hepatocyte-binding domain is conformationally dependent. To further characterize the primary structure of the hepatocyte-binding domain on the pre-S protein, a phage-displayed 15-mer peptide library and a 8-mer solid phase peptide library were used to analyze the fine specificity of the monoclonal antibody MA 18/7. Several mimotopes were identified with the phage-displayed peptide library, the majority of which possess a central motif with at least three identical residues present within the native pre-S1 sequence. No significant consensus sequences were found when these mimotopes were compared to the pre-S2 sequence. Mimotopes identified using the solid-phase peptide library also contained a similar motif. All phage mimotopes and a single mimotope from the solid-phase peptide library competed with recombinant HBsAg particles containing the pre-S1 region for binding to MA 18/7. Mouse antisera raised against four mimotopes from the phage display library reacted with HBsAg particles containing pre-S sequences. The data show that the structure of the pre-S molecule around the conserved DPAF motif in the pre-S region may have a functional role in binding HBV to cellular receptors, and that the central motif identified in mimotopes of this region may offer a novel strategy target for the improvement of existing hepatitis a vaccines which, at present, are mostly devoid of pre-S specificities. (C) 1997 Academic Press.