PROTEIN DISSECTION OF THE ENVELOPE (GP120/GP41) OF HIV

（改编自申请）广泛的长期目标 这里提出的研究是为了增加有关结构/功能的知识 HIV 包膜糖蛋白复合物 (gp120/gp41) 的关系。在 尽管付出了巨大的努力，但仍未能获得 gp120、gp41 或 gp120/gp41 复合物的详细结构。认识 这段历史，解决问题的新方法，利用蛋白质解剖， 结合噬菌体展示和组合化学，将应用于 拟议的研究。 拟议研究的三个具体目标是： 1. 确定高分辨率X射线晶体结构 HIV gp41 的三聚体核心。 2. 通过以下方法鉴定 gp120/gp41 复合物的稳定、折叠子片段 蛋白质解剖，获得这些的衍射质量晶体 子片段，并通过 X 射线解析这些子片段的结构 晶体学方法。 3. 要识别使 gp120 结构刚性化的配体，请使用这些 配体以获得 gp120 的衍射质量晶体并解决 通过 X 射线晶体学分析这些配体-gp120 配合物的结构 方法。 4. 鉴定 HIV 感染的 D 肽抑制剂，并表征 这些抑制剂的结构与 gp41 片段结合。 病毒颗粒需要 HIV 的包膜糖蛋白 (Env gp) 附着于细胞并随后病毒和细胞融合 膜。因此，包膜对于细胞的感染至关重要 艾滋病毒。此外，基本上所有的中和抗体活性 HIV-1感染的宿主针对Env gp。对于这些和其他 原因，确定 gp120-gp41 复合物的结构，以及 其组成部分，一直是努力开发新产品的主要目标 治疗艾滋病的方法。尽管经过无数努力，高 缺乏有关 Enz gp 的分辨率结构信息。现在的 提案旨在通过新方法获取这些信息。 这项研究旨在加深我们对 HIV包膜的结构/功能关系最终 促进治疗艾滋病的基于结构的药物设计。

(Adapted from the application) The broad, long-term objective of the research proposed here is to increase knowledge about structure/function relationships for the HIV envelope glycoprotein complex (gp120/gp41). In spite of considerable efforts, it has not yet been possible to obtain a detailed structure for gp120, gp41 or the gp120/gp41 complex. Recognizing this history, new approaches to the problem, utilizing protein dissection, coupled with phage display and combinatorial chemistry, will be applied in the proposed research. The three specific aims of the proposed research are: 1. To determine the high resolution X-ray crystal structure of the trimeric core of HIV gp41. 2. To identify stable, folded subfragments of the gp120/gp41 complex by protein dissection, obtain diffraction-quality crystals of these subfragments, and solve the structures of these subfragments by X-ray crystallographic methods. 3. To identify ligands that rigidify the structure of gp120, use these ligands to obtain diffraction-quality crystals of gp120 and solve the structures of these ligand-gp120 complexes by X-ray crystallographic methods. 4. To identify D-peptide inhibitors of HIV infection, and characterize the structures of these inhibitors bound to fragments of gp41. The envelope glycoprotein (Env gp) of HIV is required for virion attachment to cells and subsequent fusion of the viral and cellular membranes. The envelope is therefore crucial for infection of cells by HIV. In addition, essentially all of the neutralizing antibody activity in HIV-1 infected hosts is directed against Env gp. For these and other reasons, determining the structure of the gp120-gp41 complex, and components thereof, has been a major goal of efforts to develop new approaches for the treatment of AIDS. In spite of numerous efforts, high- resolution structural information about Enz gp is lacking. The present proposal is aimed at obtaining this information through new approaches. This research is designed to advance our understanding of structure/function relationships of the HIV envelope, ultimately facilitating structure-based drug design for the treatment of AIDS.