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

(改编自申请) 这里提出的研究是为了增加关于结构/功能的知识 HIV包膜糖蛋白复合体(gp120/gp41)的关系。在……里面 尽管作出了相当大的努力，但还不可能获得一份 Gp120、gp41或gp120/gp41复合体的详细结构。认识 这段历史，解决这个问题的新方法，利用蛋白质解剖， 与噬菌体展示和组合化学相结合，将在 建议的研究。 拟议研究的三个具体目标是： 1.确定高分辨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复合体的稳定折叠亚片段 蛋白质分离，获得这些晶体的衍射级晶体 亚片段，并用X射线解析这些亚片段的结构 结晶学方法。 3.要确定使gp120结构僵化的配体，请使用这些 配体以获得衍射级的gp120晶体并解决 这些配体-gp120配合物的X-射线结晶学结构 方法：研究方法。 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。 HIV包膜糖蛋白(Env Gp)是病毒粒子所必需的 附着在细胞上以及随后病毒和细胞的融合 膜。因此，包膜对于感染细胞是至关重要的 爱滋病毒。此外，基本上所有的中和抗体活性都在 HIV-1感染的宿主是针对Env GP的。对于这些和其他 原因，确定gp120-gp41复合体的结构，以及 其组成部分，一直是努力开发新的 艾滋病的治疗方法。尽管做出了许多努力，但高- 缺乏有关Enz GP的解析结构信息。现在 该提案的目的是通过新的方法获得这一信息。 这项研究旨在增进我们对 HIV包膜的结构/功能关系，最终 促进以结构为基础的艾滋病治疗药物设计。

(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.