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）：拟议研究的长期目标是设计疫苗和小分子抑制剂来预防艾滋病毒感染的发生。病毒包膜糖蛋白 gp120 通过与宿主细胞表面的 CD4 糖蛋白和趋化因子受体相互作用，促进人类免疫缺陷病毒 (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，有助于病毒逃避人类防御系统。即使是设计用于阻断受体与 gp120 结合的抗体，也会诱导 gp120 的构象变化，从而使病毒能够不受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将病毒 gp120 糖蛋白缩小为微型蛋白系统的策略。 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被转移到结构相容的小支架上。这将重现 gp120 不连续表位的天然样构象和结合​​功能。这种工程化的微型蛋白代表了一种独特的工具，可以在 HIV 进入宿主细胞的复杂过程中在分子水平上研究 gp120 与抗体、CD4 和趋化因子受体的相互作用。因此，该信息可用于设计和测试针对病毒进入宿主细胞的疗法。多表位方法将表位限制为其天然构象，并确保诱导针对多个抗原靶标的免疫，并且与仅使用表位（合成肽）方法相比，更有可能产生保护性免疫。为了实现这些目标；我们为这项拨款提案设定了以下具体目标： 1) 制造含有在树眼镜蛇毒素提供的分子支架内表达的 HIV gp120 表位的重组微型蛋白模拟物。 2)通过NMR波谱法评估转移至小蛋白系统的gpl20表位的构象性质。 3) 测试工程微蛋白与CD4和趋化因子受体结合的能力，以及它们引发抗体的能力。这种工程化 mirti 蛋白模拟物的方法也可能在探索其他复杂的生物过程中有用，并且也可用于测试其他疾病的新疗法。