ACTIVE CONFORMATION OF LAMININ PEPTIDE 11

67 kDa高亲和力层粘连蛋白结合蛋白（LBP）是细胞 表面蛋白，其表达与 在许多实体瘤中进展。 这种相关性可能是由于 蛋白质介导蛋白质与蛋白质之间的高亲和力相互作用的能力， 转移性肿瘤细胞与基底膜层粘连蛋白分子。 LBP的主要配体结合结构域存在于层粘连蛋白-1中， B链在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 实验一直受到可用LBP量低的限制。 研究人员不能从他们的中国人身上生产足够的蛋白质。 用于常规NMR的仓鼠卵巢（CHO）过表达系统 实验，并在目前的建议，他们计划完成核磁共振 对与LBP结合的肽11的结构研究。 调查人员 还将对有前途的环状类似物进行结构研究， 肽11。 肽本身在细胞中的寿命非常短， 血流和适度的亲和力，而非肽结构 预期模拟物具有显著改善的药理学活性， 特色 这种模拟物可以提供非常有用的佐剂 恶性实体瘤的治疗。 工作的长远目标 是使用肽11的活性构象，当它结合到 67 kDa LBP，作为基于结构设计的新型抗- 转移性药物 然而，这需要发展一个非常 准确的模板结构， 应用会产生。 调查人员还将进行一项 一系列的实验来增加我们的生物化学知识 和LBP的膜结合形式的作用机制。 这些实验将使用我们的基于肽11的光标记探针 为了便于鉴定与LBP相互作用的LBP残基， 肽11。 其他实验将评估假设，即67 kDa LBP可能与a6 b1整合素在组织中协同作用。 入侵

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.