Rational Engineering of HIV gp 120 Miniprotein Mimetics

HIV gp 120 微蛋白模拟物的合理工程

• 批准号: 6696437
• 负责人: MAHESH JASEJA
• 金额: $ 13.08万
• 依托单位: NORTH DAKOTA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-08-01 至 2007-01-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6696437
• 关键词: AIDS therapy; AIDS vaccines; HIV envelope protein gp120; HIV infections; antiAIDS agent; antigen antibody reaction; biomimetics; biotechnology; chimeric proteins; conformation; cytokine receptors; enzyme linked immunosorbent assay; monoclonal antibody; nuclear magnetic resonance spectroscopy; protein engineering; protein structure function; recombinant proteins; reptile poison; synthetic peptide; vaccine development

DESCRIPTION (provided by applicant): The long-term goal of the proposed research is to design vaccines and small molecule inhibitors to prevent the onset of HIV infection. The viral envelope glycoprotein gp120 facilitates the entry of human immunodeficiency virus (HIV) into host cells by interacting with the CD4 glycoprotein and a chemokine receptor on the host cell surface. The native structure of gp120 and its ability to undergo conformational changes during such events, help the virus to evade the human defense system. Even the antibodies designed to block receptor binding to gp120, induce conformational changes in gp120, hence enabling the virus to enter host cells with impunity. Our rationale for designing effective protective HIV vaccine and small molecule fusion inhibitors is based on a strategy to downsize the viral gp120 glycoprotein to a mini-protein system. Multiple discontinuous functional elements of gp120 antigenic CD4 and chemokine receptor binding sites, will be transferred to a structurally compatible small scaffold. This will reproduce native-like conformation and binding function of the gp120 discontinuous epitopes. Such engineered mini-proteins represent a unique tool to study the interaction of gp120 with antibodies, CD4 and chemokine receptors at the molecular level in the complex process of HIV entry into the host cells. This information thus would be useful in designing and testing therapies against viral entry in host cells. The multi-epitope approach constrains the epitopes to their native conformations, and ensures the induction of immunity against multiple antigenic targets, and would be more likely to produce protective immunity compared to using epitope only (synthetic peptide) approach. In order to accomplish these objectives; we have set the following specific aims for this grant proposal: 1) To make recombinant miniprotein mimetics containing HIV gp120 epitopes expressed within the molecular scaffold provided by dendroaspin. 2) To evaluate the conformational nature of the gpl20 epitopes transferred to the miniprotein system by NMR spectroscopy. 3) To test the ability of the engineered miniproteins to bind to CD4 and chemokine receptors, and their ability to elicit antibodies. This approach of engineered mirtiprotein mimeties may also have utility in probing other complex biological processes and a means to test newer therapies for other diseases as well.

描述（由申请人提供）：拟议的研究的长期目标是设计疫苗和小分子抑制剂，以防止HIV感染发作。病毒包膜糖蛋白GP120通过与CD4糖蛋白与宿主细胞表面上的CD4糖蛋白和趋化因子受体相互作用，促进了人类免疫缺陷病毒（HIV）进入宿主细胞。 GP120的天然结构及其在此类事件中发生构象变化的能力，可以帮助病毒逃避人类的防御系统。即使是旨在阻断受体与GP120结合的抗体，也会诱导GP120的构象变化，从而使该病毒能够不惩罚地进入宿主细胞。 我们设计有效的保护性HIV疫苗和小分子融合抑制剂的基本原理是基于一种策略，将病毒GP120糖蛋白缩小为微型蛋白质系统。 GP120抗原CD4和趋化因子受体结合位点的多个不连续功能元件将转移到结构兼容的小支架上。这将重现GP120不连续表位的天然样构象和结合​​功能。这种工程化的迷你蛋白代表了研究GP120与分子水平的抗体，CD4和趋化因子受体相互作用的独特工具，在HIV进入宿主细胞的复杂过程中。因此，这些信息对于针对宿主细胞中的病毒进入的设计和测试疗法将很有用。多诊断方法将表位限制在其天然构象上，并确保免疫对多种抗原靶标的诱导，并且与仅使用表位（合成肽）方法相比，更可能产生保护性免疫。为了实现这些目标；我们设定了该赠款提案的以下特定目的：1）使重组小蛋白模仿含有HIV GP120表位，该表达在Dendroaspin提供的分子支架中表达。 2）评估NMR光谱传递到微蛋白系统的GPL20表位的构象性质。 3）测试工程小动物与CD4和趋化因子受体结合的能力，以及它们引起抗体的能力。这种工程的mirtiprotein Mimeties的方法也可能具有效用，可以探索其他复杂的生物学过程，也可以测试其他疾病的新疗法。