Determining structural features of a collagen lysyl hydroxylase that promotes lung cancer metastasis

项目摘要/摘要： 应聘者：郭博士接受过生物化学和结构生物学方面的广泛培训。他是一名博士后 MD Anderson癌症中心研究员试图了解胶原赖氨酸羟基酶(LH)的结构 特征调节肺癌的转移。郭博士是一位高效率的研究员，有18位同行评议 论文(其中6篇为第一作者或共同第一作者)。作为对他成就的认可，他收到了无数 在他的博士和博士后培训期间获奖。 职业发展/培训：郭博士在MD Anderson的导师包括乔纳森·库里博士，一位内科医生- 在肺癌生物学和人类肺癌小鼠模型方面有专长的科学家，以及约翰·泰纳博士， 一位著名的蛋白质结晶学家，研究大分子组装以指导药物设计。另外， 郭博士将接受三尾山内博士(北卡罗来纳大学教堂山)的培训，他是一名胶原生物化学家，曾 这是最早阐明LHS如何修饰胶原蛋白的人之一。这些调查人员设计了一项培训 以他们的关键科学学科为中心的计划，将增强郭博士在各自的 并提供平稳过渡到独立所需的技能。 研究：我们的研究小组已经表明，LH2的高表达会推动肺癌的转移，并诱导 肿瘤间质中的胶原交联型开关。LH2是一个值得关注的治疗靶点，但具有选择性的促黄体生成素抑制剂 这是一个缺陷，部分原因是缺乏对LH2的结构性洞察。我的提议的目的是 目的是阐明促进肺癌转移的LH2结构特征。在我的初步结果中，我 描述生产人LH2蛋白的新方法，并测定其活性和催化结构 发现一个同源二聚体由Fe+2结合稳定。我发现两个人 活性位点位于二聚体界面深表面裂缝的两侧，这表明二聚化作用产生了一种胶原- 结合部位，与活性部位相邻碱性残基定位形成盐桥 胶原蛋白上的端肽酸性残基。根据这些发现，我推测LH2具有端肽-1H。 由于独特的特性，它可以形成与Fe2+相互作用的稳定的二聚体结构 胶原蛋白上的端肽赖氨酸。我将通过确定LH二聚体组装如何来检验这一假设 由Fe2+结合稳定并与端肽赖氨酸残基相互作用调节胶原交联物的形成 和肺肿瘤的发生。 总括而言，我的建议会针对肺癌转移的临床问题。新奇之处在于 初步结果提供了对LH胶原蛋白的第一个结构洞察，对当前的假设提出了挑战 促进黄体生成素酶活性量化和候选评估的范例和技术 肿瘤转移的驱动力。

Project Summary/Abstract: Candidate: Dr. Guo has received broad training in biochemistry and structural biology. He is a postdoctoral fellow at MD Anderson Cancer Center seeking to understand how collagen lysyl hydroxylase (LH) structural features regulate lung cancer metastasis. Dr. Guo is a highly productive investigator, with 18 peer-reviewed papers (6 of which as first or co-first author). As recognition of his achievements, he received numerous awards during his doctoral and postdoctoral training. Career Development/Training: Dr. Guo's mentors at MD Anderson include Dr. Jonathan Kurie, a physician- scientist with expertise in lung cancer biology and mouse modeling of human lung cancer, and Dr. John Tainer, a renowned protein crystallographer studying macromolecular assemblies to inform drug design. Additionally, Dr. Guo will receive training from Dr. Mitsuo Yamauchi (UNC-Chapel Hill), a collagen biochemist who was among the first to elucidate how collagens are modified by LHs. These investigators have designed a training program centered on their key scientific disciplines that will strengthen Dr. Guo's abilities in their respective fields and provide the skills needed for a smooth transition to independence. Research: Our group has shown that high expression of LH2 drives lung cancer metastasis and induces a collagen cross-link switch in tumor stroma. LH2 is a therapeutic target of interest, but selective LH inhibitors are not available, a deficiency due in part to a lack of structural insight into LH2. The objective of my proposal is to elucidate LH2 structural features that promote lung cancer metastasis. In my preliminary results, I describe new methods to produce human LH2 protein and assay its activity and the structure of the catalytic domain of a viral LH homologue, which revealed a homodimer stabilized by Fe+2-binding. I found that the two active sites flank a deep surface cleft on the dimer interface, suggesting that dimerization creates a collagen- binding site, and that basic residues adjacent to the active site are positioned to form salt bridges with telopeptidyl acidic residues on collagen. From these findings, I hypothesize that LH2 has telopeptidyl-LH activity owing to unique features that allow it to form Fe2+-stabilized dimeric structures that interact with telopeptidyl lysines on collagen. I will test this hypothesis by determining how LH dimer assemblies are stabilized by Fe2+-binding and interact with telopeptidyl lysine residues to regulate collagen cross-link formation and lung tumorigenesis. In sum, my proposal will address the clinical problem of lung cancer metastasis. The novelty rests in preliminary results that provide the first structural insights into a collagen LH, hypotheses that challenge current paradigms, and technologies that facilitate quantification of LH enzymatic activity and evaluation of candidate metastasis drivers.