Fontenot J D, Zacharopoulos V R, Phillips D M
The Population Council, New York, New York 10021, USA.
J Virol. 1996 Oct;70(10):6557-62. doi: 10.1128/JVI.70.10.6557-6562.1996.
The proline-rich tandem repeat domain of human mucin MUC1 forms an extended structure containing large repeating loops that are crested by a turn. We show that the repeating-loop structure of MUC1 can be replaced by an antibody complementarity-determining region loop of a human immunodeficiency virus type 1 (HIV-1)-specific neutralizing antibody to create a chimeric, multivalent, mucin-like, anti-HIV-1 compound. We used 8 residues of an antibody molecule to replace 8 of 20 residues of the MUC1 tandem-repeat sequence. The antiviral peptide discussed here contains three copies of a 20-residue tandem repeat, (IYYDYEEDPAPGSTAPPAHG)3, for a total of 60 residues. We demonstrate that the mucin-antibody chimera retains the binding specificity of the parent antibody (monoclonal antibody F58), GPGR of the HIV-1 gp120 V3 neutralizing epitope, and the ability to neutralize virus particles. In inhibition enzyme-linked immunosorbent assay, the mucin-antibody chimeric peptide could inhibit 71 to 84% of binding to a V3 loop peptide by monoclonal antibodies known to be specific for GPGR in the V3 loop. The mucin-antibody chimeric peptide could also inhibit monoclonal antibody binding to native gp120 captured from virus particles. In addition, the chimeric peptide neutralized the homologous HIV-IIIB virus in a standard neutralization assay. The methods of antiviral peptide design and construction presented here are general and theoretically limited only by the size of the antibody repertoire. This approach could be used to synthesize peptides for a variety of therapeutic applications.
人黏蛋白MUC1富含脯氨酸的串联重复结构域形成一种延伸结构,其中包含由一个转角加帽的大重复环。我们发现,MUC1的重复环结构可被人免疫缺陷病毒1型(HIV-1)特异性中和抗体的抗体互补决定区环取代,从而创建一种嵌合、多价、黏蛋白样抗HIV-1化合物。我们用抗体分子的8个残基取代了MUC1串联重复序列20个残基中的8个。此处讨论的抗病毒肽包含20个残基串联重复序列的三个拷贝,即(IYYDYEEDPAPGSTAPPAHG)3,总共60个残基。我们证明,黏蛋白-抗体嵌合体保留了亲本抗体(单克隆抗体F58)的结合特异性、HIV-1 gp120 V3中和表位的GPGR以及中和病毒颗粒的能力。在抑制酶联免疫吸附测定中,黏蛋白-抗体嵌合肽可抑制已知对V3环中GPGR具有特异性的单克隆抗体与V3环肽结合的71%至84%。黏蛋白-抗体嵌合肽还可抑制单克隆抗体与从病毒颗粒捕获的天然gp120的结合。此外,在标准中和试验中,嵌合肽中和了同源的HIV-IIIB病毒。本文介绍的抗病毒肽设计和构建方法具有通用性,理论上仅受抗体库大小的限制。这种方法可用于合成多种治疗应用的肽。