权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

MULTI-DIMENSIONAL NMR OF HIV-1 ENVELOPE GLYCOPROTEINS

HIV-1 包膜糖蛋白的多维核磁共振

基本信息

批准号：
7917499
负责人：
JACOB ANGLISTER
金额：
$ 19.52万
依托单位：
WEIZMANN INSTITUTE OF SCIENCE
依托单位国家：
美国
项目类别：
财政年份：
1997
资助国家：
美国
起止时间：
1997-09-01 至 2012-08-31
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): The long term goals of our research are to learn about the mechanism for HIV-1 target recognition, to understand the selectivity of the virus to different cells of the immune system and to study mechanisms of viral entry inhibition. The binding of human immunodeficiency-virus type-1 (HIV-1) to its target cells is mediated primarily by the envelope glycoprotein (gp120) of the virus. Initially gp120 binds to CD4, a molecule found on the surface of both T-cells and macrophages. This binding triggers a conformational change in gp120 that forms another binding site to either CCR5 or CXCR4 chemokine receptors, which serve as co-receptors for HIV-1 binding. After binding to the co-receptor, gp120 undergoes another conformational change that exposes the gp41 trans-membrane glycoprotein of the virus which mediates HIV- 1 fusion with its target cell membrane. The peptide T-20 (Fuzeon), used as a viral entry inhibitor for the treatment of HIV-1 infected individuals, corresponds in its sequence to the C-terminal half of the gp41 region HR2 and a segment down stream of HR2. The structure of the co-receptor binding site on gp120 is still elusive and the mechanism for co-receptor selectivity is still unknown. Similarly, for T-20, the mechanism by which it inhibits fusion is not understood. Particularly enigmatic is the role played by its nine C-terminal residues, without which this anti-HIV drug loses its potency in entry-inhibition. The specific aims of the current proposal are as follows: a) Learn how the flexibility of gp120 enables the formation of the binding site for CD4 and CCR5, b) Learn how HIV-1 co-receptor selectivity is determined, and c) Investigate the interactions of T-20 with possible targets on gp41 and gp120 and reveal the role of the C-terminal segment T-20 in viral inhibition. We will use a novel multi-dimensional NMR technique designed to solve the structure of large proteins without requiring deuteration. The solution structure of HIV-1 gp120 in the unliganded form, in complex with a CD4-mimic peptide (miCD4) and in ternary complex with miCD4 and CCR5 extra-cellular fragments will be determined. In addition, we will determine the structure of the anti-HIV-1 drug T-20 in complex with gp41 and gp120 targets. When solved, the solution structure of HIV-1 gp120 will be probably the first structure of a protein expressed and labeled in mammalian cells and one of the largest structures solved by NMR. Considering the flexibility of gp120 that is directly related to its function, structure determination in solution is of utmost importance. The structure of gp120 in its three different conformations will enable us to learn about the mechanism for CD4 and co-receptor recognition and for designing anti-HIV- 1 immunogens, antagonists and entry inhibitors that target the gp120 and CCR5 binding sites. Studies of T- 20 complexes with gp41 and gp120 segments will reveal the role of the C-terminal segment of T-20 in viral inhibition and contribute to the design more potent entry inhibitors.

描述(申请人提供)：我们研究的长期目标是了解HIV-1靶标识别的机制，了解病毒对免疫系统不同细胞的选择性，并研究病毒进入抑制的机制。人类免疫缺陷病毒1型(HIV-1)与其靶细胞的结合主要是由病毒的包膜糖蛋白(Gp120)介导的。最初，gp120与T细胞和巨噬细胞表面的一种分子CD4结合。这种结合触发了gp120的构象变化，形成了与CCR5或CXCR4趋化因子受体的另一个结合部位，这两种受体是HIV-1结合的辅助受体。在与辅助受体结合后，gp120经历了另一次构象变化，暴露了病毒的gp41跨膜糖蛋白，它介导了HIV-1与其靶细胞膜的融合。T-20(Fuzeon)是一种病毒进入抑制剂，用于治疗HIV-1感染者，其序列对应于gp41区域HR2的C末端一半和HR2下游的一段。Gp120上共受体结合部位的结构仍不清楚，共受体选择性的机制仍不清楚。同样，对于T-20，它抑制融合的机制尚不清楚。尤其令人费解的是它的九个C末端残基所起的作用，如果没有这些残基，这种抗艾滋病毒药物就会失去进入抑制的效力。目前建议的具体目标如下：a)了解gp120的灵活性如何使CD4和CCR5结合位点的形成，b)了解HIV-1共同受体的选择性是如何决定的，以及c)研究T-20与gp41和gp120上可能的靶点的相互作用，并揭示C末端片段T-20在病毒抑制中的作用。我们将使用一种新的多维核磁共振技术来解决大蛋白质的结构问题，而不需要重氢。将确定HIV-1gp120的未连接形式、与CD4模拟肽(MiCD4)的复合体以及与miCD4和CCR5胞外片段的三元复合体的结构。此外，我们还将确定抗HIV-1药物T-20与gp41和gp120靶点的复合体的结构。当解决时，HIV-1gp120的溶液结构可能是在哺乳动物细胞中表达和标记的第一个蛋白质结构，也是通过核磁共振解析的最大结构之一。考虑到gp120与其功能直接相关的灵活性，溶液中结构的确定是至关重要的。Gp120三种不同构象的结构将使我们能够了解CD4和辅助受体识别的机制，以及设计针对gp120和CCR5结合位点的抗HIV-1免疫原、拮抗剂和进入抑制剂。对T-20与gp41和gp120片段的复合体的研究将揭示T-20的C末端片段在病毒抑制中的作用，并有助于设计更有效的进入抑制剂。

项目成果

期刊论文数量（28）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

An optimally constrained V3 peptide is a better immunogen than its linear homolog or HIV-1 gp120.

DOI：
10.1016/j.virol.2010.03.007
发表时间：
2010-06-05
期刊：
Virology
影响因子：
3.7
作者：
Moseri A;Tantry S;Sagi Y;Arshava B;Naider F;Anglister J
通讯作者：
Anglister J

Intermolecular interactions in a 44 kDa interferon-receptor complex detected by asymmetric reverse-protonation and two-dimensional NOESY.

DOI：
10.1021/bi100041f
发表时间：
2010-06-29
期刊：
Biochemistry
影响因子：
2.9
作者：
Nudelman I;Akabayov SR;Schnur E;Biron Z;Levy R;Xu Y;Yang D;Anglister J
通讯作者：
Anglister J

HIV-1 peptide vaccine candidates: selecting constrained V3 peptides with highest affinity to antibody 447-52D.

HIV-1肽疫苗候选物：选择与抗体447-52D具有最高亲和力的受限V3肽。

DOI：
10.1021/bi900146g
发表时间：
2009
期刊：
Biochemistry
影响因子：
2.9
作者：
Mester,Brenda;Manor,Revital;Mor,Amit;Arshava,Boris;Rosen,Osnat;Ding,Fa-Xiang;Naider,Fred;Anglister,Jacob
通讯作者：
Anglister,Jacob

A model of a gp120 V3 peptide in complex with an HIV-neutralizing antibody based on NMR and mutant cycle-derived constraints.

基于 NMR 和突变体循环衍生约束的 gp120 V3 肽与 HIV 中和抗体复合物的模型。