NMR Studies of Plasminogen and Related Genomic Domains

纤溶酶原及相关基因组结构域的 NMR 研究

• 批准号: 6638226
• 负责人: MIGUEL LLINAS
• 金额: $ 37.19万
• 依托单位: CARNEGIE-MELLON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1982
• 资助国家: 美国
• 起止时间: 1982-07-01 至 2005-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6638226
• 关键词: agrin; angiostatins; chemical association; chemical binding; collagenase; conformation; endopeptidases; extracellular matrix; fibronectins; gelatin; genome; intermolecular interaction; kininogens; molecular cloning; molecular dynamics; nuclear magnetic resonance spectroscopy; plasminogen; plasminogen activator; plasminogen activator inhibitors; protein structure function; protein tyrosine kinase; recombinant proteins; site directed mutagenesis; thrombospondins; urokinase

DESCRIPTION: (Investigator's abstract) We propose to investigate via NMR spectroscopy the structure and function of genomic protein modules and multimodular arrays related to plasminogen (Pgn) and its activators urokinase (uPA) and tissue Pgnactivator (tPA). These proteins are found both in blood plasma and in the extracellular matrix (ECM) where they play crucial roles in the fibrinolytic dissolution of blood clots, cell proliferation and migration, embryogenesis, tissue remodeling metastasis, etc. Pgn, tPA and uPA contain kringle (K) domains that mediate their binding to specific substrates. The Pgn kringles interact with the inhibitor a2-antiplasmin C- and the Pgn N-terminal peptide domains. Interestingly, the Pgn KI, K2, K3, K5, their tandem arrays K123, K1234 (angiostatin), and K12345, as well as the uPA peptide KPSSPPEE 143 inhibit endothelial cell growth and migration, thus tissue and tumor vascularization. Antiangiogenic activities also are displayed by the blood plasma/ECM proteins kininogen D5 (a Zn2+-binding domain) and thrombospondin 2nd type-1 domain. Implicated in tissue remodeling is the matrix metalloproteinase 2, which digests denatured collagen (gelatin) via adhesion through three fibronectin type II domains. Activation of Pgn by uPA, a key step in metastatic cell propagation, involves a membrane-anchored receptor (uPAR) which contains three snake neurotoxin-type modules. Related to uPA activity is the receptor associated protein (RAP), a chaperon that stabilizes newly synthesized low density lipoprotein receptor-related protein and the very low density lipoprotein receptor, presumably via its C-terminal domain (ctRAP). Pgn and tPA also are found in brain where their presence correlates with memory processes. In addition, in brain is neurotrypsin, a novel kringle containing proteinase. Of related neurological interest is the SEA module of agrin, a protein produced by motoneurons that induces the aggregation of nicotinic acetylcholine receptors. Functional in nerve tissue extension are various transmembrane protein tyrosine kinase receptors which contain kringle and frizzled (cysteine-rich) domains, likely to be involved in ligand binding. A main thrust of the project will be the development of CLOUDS, a relaxation matrix approach that avoids the assignment bottleneck and aims at high throughput structural analysis of protein NMR data.

描述：（研究者摘要）我们建议通过 NMR 进行研究 光谱学基因组蛋白质模块的结构和功能以及 与纤溶酶原 (Pgn) 及其激活剂尿激酶相关的多模块阵列 (uPA) 和组织 Pgnactivator (tPA)。这些蛋白质存在于血液中 血浆和细胞外基质 (ECM) 中它们发挥着至关重要的作用 血凝块的纤溶溶解、细胞增殖和迁移， 胚胎发生、组织重塑、转移等。Pgn、tPA和uPA含有 kringle (K) 结构域介导其与特定底物的结合。 PGN kringles 与抑制剂 a2-抗纤溶酶 C 端和 Pgn N 端相互作用 肽结构域。有趣的是，Pgn KI、K2、K3、K5，它们的串联阵列 K123、K1234（血管抑制素）和 K12345，以及 uPA 肽 KPSSPPEE 143 抑制内皮细胞的生长和迁移，从而抑制组织和肿瘤 血管化。血液也显示出抗血管生成活性 血浆/ECM 蛋白激肽原 D5（Zn2+ 结合结构域）和血小板反应蛋白 2nd 1 类域。基质金属蛋白酶参与组织重塑 2、通过三个粘连消化变性的胶原蛋白（明胶） 纤连蛋白 II 型结构域。 uPA 激活 Pgn，这是转移的关键步骤 细胞增殖涉及膜锚定受体（uPAR），其中包含 三个蛇神经毒素型模块。与uPA活性相关的是受体 相关蛋白（RAP），一种稳定新合成的低分子伴侣 密度脂蛋白受体相关蛋白和极低密度 脂蛋白受体，可能是通过其 C 末端结构域 (ctRAP)。 Pgn 和 tPA 也存在于大脑中，它们的存在与记忆过程相关。 此外，大脑中还有神经胰蛋白酶，一种含有蛋白酶的新型三环蛋白。 相关的神经学兴趣是 agrin 的 SEA 模块，agrin 是一种产生的蛋白质 通过诱导烟碱乙酰胆碱聚集的运动神经元 受体。在神经组织延伸中发挥功能的是各种跨膜 含有 kringle 和 frizzled 的蛋白酪氨酸激酶受体 （富含半胱氨酸）结构域，可能参与配体结合。一个主攻 该项目的重点将是开发 CLOUDS，一种松弛矩阵方法 避免分配瓶颈并以高吞吐量结构为目标 蛋白质核磁共振数据分析。