PHYSICAL INTERACTIONS OF HIV

HIV 的身体相互作用

• 批准号: 3145401
• 负责人: BRADFORD A JAMESON
• 金额: $ 16.27万
• 依托单位: THOMAS JEFFERSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-07-01 至 1993-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3145401
• 关键词: AIDS; CD4 molecule; DNA footprinting; antigen antibody reaction; antiviral antibody; human immunodeficiency virus; human immunodeficiency virus 1; laboratory rabbit; peptide chemical synthesis; protein sequence; protein structure function; receptor binding; tissue /cell culture; virus protein

The Cd4 protein serves as the major protein receptor for the human immunodeficiency virus (HIV). Expression constructs of the first 104 amino acids of the protein (the V1 domain) can bind with high affinity to the gp120 chain variable domain and has been used to classify the CD4 protein as a member of the Ig-superfamily of proteins. Mutational data suggests that a small region within the V1 domain of the CD4 protein (residues 38- 55) is responsible for the observed binding of this domain to the gp120 protein. The region 38-55 covers a subdomain of the CD4 protein that corresponds to one of the three antigen-antibody contact structures within the light chain variable domain (the three antibody contact domains are referred to as CDR1, CDR2 and CDR3). Our preliminary results provide the first direct evidence that the CDR2-like domain of the CD4 protein (residues 45-54)are directly involved in contacting the gp120 protein. Furthermore, we have evidence that the CDR3 domain (CD4 residues 84-101) is also involved in this contact as well as suggestive evidence of the involvement of the CDR1-like domain. Using a known light chain variable domain structure as a folding template, we have modeled the V1 domain of the CD4 protein. We have used our CD4 model as a structural template for the design of conformationally-restricted synthetic peptides. Our preliminary work with both the CDR2-like domain of the CDR2- and CDR3-like domains of the CD4 protein were performed using these restricted analogs. We have shown that peptides derived from both of these restricted analogs. We have shown that peptides derived from both of the CDR-like domains are capable of potently inhibiting virus binding to CD4 positive cells. Here we propose to expand our use of the conformationally-restricted analogs to identify CD4/gp120 contact regions as well as refine such structures to yield a panel of high affinity reagents capable of preventing the HIV-CD4 interaction. Based on our preliminary results and the known structure/function relationships within the Ig-superfamily of proteins, we propose that the gp120 contact surface on the CD4 protein is directly analogous to the antigen contact surface on an antibody. Finally, in order to prove (or disprove) and extend this hypothesis, we propose to perform protein footprinting experiments on the CD4/gp120 complex. This will identify protein regions sequestered when the protein-protein complex is formed.

CD4蛋白是人类的主要蛋白受体 免疫缺陷病毒（HIV）。 第一个104氨基的表达构造 蛋白质（V1结构域）的酸可以与高亲和力结合 GP120链可变域，已用于对CD4蛋白进行分类 作为蛋白质Ig-Superfaily的成员。 突变数据建议 在CD4蛋白的V1结构域中的一个小区域（残基38-- 55）负责观察到该域与GP120的结合 蛋白质。 区域38-55涵盖了CD4蛋白的子域 对应于三个抗原抗体接触结构之一 轻链可变域（三个抗体接触域是 称为CDR1，CDR2和CDR3）。 我们的初步结果提供了 第一个直接证据表明CD4蛋白的CDR2样结构域 （残基45-54）直接参与接触GP120蛋白。 此外，我们有证据表明CDR3域（CD4残基84-101）是 还参与了这种联系以及暗示性证据 CDR1状域的参与。 使用已知的轻链变量 域结构作为折叠模板，我们已经建模了 CD4蛋白。 我们已经使用CD4模型作为结构模板 构型限制的合成肽的设计。 我们的 与CDR2和CDR3一样的CDR2状域的初步工作 使用这些限制的类似物进行CD4蛋白的结构域。 我们已经表明，从这两个限制的类似物中得出的肽。 我们已经表明，从两个CDR样域中得出的肽是 能够有效抑制病毒与CD4阳性细胞结合。 这里 我们建议将对构象限制类似物的使用扩展到 识别CD4/GP120接触区域，并将此类结构改进到 产生一组能够防止HIV-CD4的高亲和力试剂 相互作用。 根据我们的初步结果和已知的 蛋白质的Ig-Superfamily中的结构/功能关系，我们 提出CD4蛋白上的GP120接触表面直接 类似于抗体上的抗原接触表面。 最后，按顺序 为了证明（或反驳）并扩展此假设，我们建议执行 CD4/GP120复合物上的蛋白质足迹实验。 这会 当蛋白质蛋白络合物是 形成。