METALLOPEPTIDES OF GP41 IN HIV-1 FUSION INHIBITION

GP41 的金属肽抑制 HIV-1 融合

• 批准号: 6846609
• 负责人: MIRIAM GOCHIN
• 金额: $ 10.84万
• 依托单位: UNIVERSITY OF THE PACIFIC-STOCKTON
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-05-01 至 2005-12-01
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6846609
• 关键词: HIV envelope protein gp41; antiAIDS agent; antiviral agents; binding sites; combinatorial chemistry; conformation; drug design /synthesis /production; fluorescent dye /probe; molecular probes; peptide analog; protein binding; protein structure function

DESCRIPTION (provided by applicant): This proposal focuses on the development of entry inhibitors against Human Immunodeficiency Virus Type-1, by targeting the viral fusion protein gp41. Gp41 mediates fusion to mammalian host cells via a highly conserved coiled coil domain. The hypothesis is that compounds that bind to this domain will act as fusion inhibitors by preventing the gp41 conformational rearrangement required for fusion. Discovery of inhibitors to the gp41 coiled coil is hampered by the fact that this domain is not readily accessible in solution, being buried in intact gp41 and unstable when excised from the rest of the protein. We have developed a stabilized coiled coil construct through the use of bidentate bipyridyl groups covalently attached at the N-termini of gp41 peptides. The addition of transition metal ion to the bipyridylated peptides stabilizes the trimeric helical structure of the coiled coil, through the formation of an octahedral tris-bipyridyl complex, and removes non-specific aggregation of the hydrophobic peptide. The goal of this proposal is to make use of the probe properties of various metal ions to establish a screening and structural assay for inhibitors which bind to the coiled coil. 1) Discrete metal ion - coordinated constructs which represent the gp41 inner coiled coil will be developed and analyzed by NMR structure determination. 2) Constructs will be used in the development and refinement of a fluorescence binding assay to detect compound binding to the coiled coil. 3) Library screening will be conducted and successful hits analyzed with in vitro syncytium and viral infectivity assays. The metal ion used in each of these specific aims will be selected to have diamagnetic, paramagnetic or fluorescent properties, as needed, to be utilized in the biophysical assays. The long term goal of this proposal is to develop promising non-peptide drug candidates for HIV-1 fusion inhibition.

描述(由申请人提供)：这项提案的重点是通过靶向病毒融合蛋白gp41开发针对人类免疫缺陷病毒1型的进入抑制剂。Gp41通过高度保守的螺旋结构域介导与哺乳动物宿主细胞的融合。假设与这个结构域结合的化合物将通过阻止融合所需的gp41构象重排而起到融合抑制作用。Gp41卷曲线圈的抑制剂的发现受到以下事实的阻碍，即该结构域在溶液中不易接触，被埋入完整的gp41中，并且当从蛋白质的其余部分中切除时不稳定。我们开发了一种稳定的卷曲结构，通过使用双齿联吡啶基团共价连接在gp41多肽的N-末端。过渡金属离子的加入通过形成八面体三联吡啶络合物稳定了卷曲螺旋的三聚体螺旋结构，并消除了疏水性多肽的非特异性聚集。该方案的目的是利用各种金属离子的探针性质，建立一种筛选和结构分析结合到盘绕线圈上的缓蚀剂的方法。1)将开发代表gp41内螺旋线圈的离散金属离子配位结构，并通过核磁共振结构测定进行分析。2)构建体将用于开发和改进荧光结合分析，以检测与盘绕线圈结合的化合物。3)进行文库筛选，用体外合胞体和病毒感染性试验分析成功的HITS。用于每个特定目的的金属离子将根据需要被选择为具有抗磁性、顺磁性或荧光性质，以用于生物物理分析。这项提议的长期目标是开发有前景的非肽类药物来抑制HIV-1融合。