ACTIVE CONFORMATION OF LAMININ PEPTIDE 11

层粘连蛋白肽 11 的活性构象

• 批准号: 2010077
• 负责人: JEAN Rosalind STARKEY
• 金额: $ 12.66万
• 依托单位: MONTANA STATE UNIVERSITY - BOZEMAN
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-04-07 至 2000-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2010077
• 关键词: CHO cells; SCID mouse; antineoplastics; binding proteins; cell line; chemical binding; conformation; drug design /synthesis /production; integrins; laboratory mouse; laminin; neoplastic growth; nuclear magnetic resonance spectroscopy; peptide analog; protein structure function; transfection

The 67 kDa high affinity laminin binding protein (LBP) is a cell surface protein whose expression shows a positive correlation with progression in many solid tumors. This correlation likely results from the ability of the protein to mediate high affinity interactions of a metastatic tumor cells with basement membrane laminin molecules. The major ligand binding domain for the LBP occurs in the laminin-1 b chain at the COPGYIGSR (peptide 11) site. The investigators have preliminary data (from NMR Tr-NOESY experiments) on the conformation of peptide 11 when bound to isolated 67 kDa LBP, but experiments have been restricted by the low amount of available LBP. The investigators can not produce sufficient protein from their chinese hamster ovary (CHO) overexpression system for routine NMR experiments, and, in the present proposal, they plan to complete NMR structural studies on peptide 11 bound to the LBP. The investigators will also pursue structural studies on promising cyclic analogs of peptide 11. The peptides, themselves, have very short lifetimes in the blood stream and modest affinity, whereas non-peptide structural mimetics are expected to have significantly improved pharmacological characteristics. Such mimetics could provide extremely useful adjuvant therapies for malignant solid tumors. The long range goal of the work is to use the active conformation of peptide 11, when it is bound to the 67 kDa LBP, as a template for structure-based design of novel anti- metastatic drugs. This, however, requires the development of a very accurate template structure which the studies outlined in the current application would produce. The investigators will also undertake a series of experiments to increase our knowledge of the biochemistry and mechanism of action of the membrane associated form of the LBP. These experiments will use our peptide 11 based photolabelling probe to facilitate identification of the LBP residues which interact with peptide 11. Other experiments will evaluate the postulate that the 67 kDa LBP may act in concert with the a6b1 integrin during tissue invasion.

67 kDa 高亲和力层粘连蛋白结合蛋白 (LBP) 是一种细胞 其表达量呈正相关的表面蛋白 许多实体瘤的进展。 这种相关性可能来自 蛋白质介导高亲和力相互作用的能力 具有基底膜层粘连蛋白分子的转移性肿瘤细胞。 LBP 的主要配体结合域出现在层粘连蛋白-1 中 b 链位于 COPGYIGSR（肽 11）位点。调查人员已 初步数据（来自 NMR Tr-NOESY 实验） 肽 11 与分离的 67 kDa LBP 结合时的构象，但是 实验受到可用 LBP 量低的限制。 研究人员无法从他们的中国人中生产出足够的蛋白质 用于常规 NMR 的仓鼠卵巢 (CHO) 过表达系统 实验，并且在目前的提案中，他们计划完成 NMR 与 LBP 结合的肽 11 的结构研究。 调查人员 还将对有前景的环状类似物进行结构研究 肽 11. 肽本身在体内的寿命非常短 血流和适度的亲和力，而非肽结构 模拟物有望显着改善药理学 特征。 这种模拟物可以提供极其有用的佐剂 恶性实体瘤的治疗。 工作的长期目标 是利用肽11的活性构象，当它与 67 kDa LBP，作为新型抗病毒药物基于结构的设计模板 转移药物。 然而，这需要开发一个非常 当前研究中概述的准确模板结构 应用程序将产生。 调查人员还将开展 一系列实验来增加我们的生物化学知识 以及 LBP 膜相关形式的作用机制。 这些实验将使用我们基于肽 11 的光标记探针 以便于鉴定与相互作用的 LBP 残基 肽 11。其他实验将评估以下假设：第 67 章 kDa LBP 可能在组织过程中与 a6b1 整合素协同作用 入侵。