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

确定促进肺癌转移的胶原赖氨酰羟化酶的结构特征

• 批准号: 10471895
• 负责人: Houfu Guo
• 金额: $ 23.68万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-11 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10471895
• 关键词: 2-Oxoglutarate 5-Dioxygenase Procollagen-Lysine; Acanthamoeba; Achievement; Active Sites; Address; Aldehydes; Alleles; Arginine; Aspartate; Award; Binding; Binding Sites; Biochemical; Biochemistry; Biological Assay; Biology; C-terminal; Cancer Biology; Cancer Center; Cartilage; Catalytic Domain; Charge; Clinical; Collagen; Complex; Crystallization; Crystallography; Development; Dimerization; Discipline; Disease; Drug Design; Enzymes; Epithelial; Epithelial Cells; Evaluation; Family member; Foundations; Future; Gelatinase B; Glutamates; Growth; Homologous Gene; Human; Hydrophobicity; Hydroxylysine; KRAS2 gene; Laboratories; Leucine; Lung; Lung Adenocarcinoma; Lung Neoplasms; Lysine; Malignant neoplasm of lung; Mentors; Metastatic Neoplasm to the Lung; Methods; Mus; Mutation; N-terminal; Neoplasm Metastasis; Paper; Peer Review; Physicians; Positioning Attribute; Postdoctoral Fellow; Proline; Proteins; Reporting; Research; Research Personnel; Rest; Scientist; Sodium Chloride; Structure; Sum; Surface; Technology; Testing; Training; Training Programs; Tumor Tissue; Tumor-infiltrating immune cells; Tyrosine; Viral; antagonist; bone; career development; clinical application; crosslink; design; dimer; human model; inhibitor; insight; interest; lung tumorigenesis; macromolecular assembly; malignant breast neoplasm; mouse model; mutant; novel; post-doctoral training; prognostic value; protein structure; sarcoma; skills; structural biology; therapeutic target; tumor; tumor growth; tumorigenesis

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.

项目摘要/摘要： 候选人：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 安德森癌症中心研究员寻求了解胶原蛋白赖氨酰羟化酶 (LH) 的结构 功能调节肺癌转移。郭博士是一位高产的研究者，拥有 18 项经过同行评审的研究成果 论文（其中第一作者或共同第一作者 6 篇）。为了表彰他的成就，他获得了无数 在博士和博士后培训期间获得奖项。 职业发展/培训：郭博士在 MD 安德森的导师包括 Jonathan Kurie 博士，他是一名医生 拥有肺癌生物学和人类肺癌小鼠模型专业知识的科学家 John Tainer 博士， 一位著名的蛋白质晶体学家，研究大分子组装以指导药物设计。此外， 郭博士将接受 Mitsuo Yamauchi 博士（北卡罗来纳大学教堂山分校）的培训，他是一位胶原蛋白生物化学家， 是最早阐明 LH 如何修饰胶原蛋白的人之一。这些研究人员设计了一个训练 项目集中于他们的关键科学学科，这将增强郭博士在各自领域的能力 领域并提供平稳过渡到独立所需的技能。 研究：我们课题组已经证明LH2的高表达会驱动肺癌转移并诱导 肿瘤基质中的胶原交联开关。 LH2 是一个感兴趣的治疗靶点，但选择性 LH 抑制剂 无法获得，这一缺陷部分是由于缺乏对 LH2 结构的了解。我的提案的目标 旨在阐明促进肺癌转移的LH2结构特征。在我的初步结果中，我 描述了生产人类 LH2 蛋白并测定其活性和催化结构的新方法 病毒 LH 同源物的结构域，揭示了通过 Fe+2 结合稳定的同二聚体。我发现两个 活性位点位于二聚体界面上深表面裂缝的侧面，表明二聚化产生了胶原蛋白- 结合位点，并且与活性位点相邻的碱性残基被定位以与形成盐桥 胶原蛋白上的端肽基酸性残基。根据这些发现，我推测 LH2 具有端肽基-LH 其独特的功能使其能够形成 Fe2+ 稳定的二聚结构，并与 胶原蛋白上的端肽基赖氨酸。我将通过确定 LH 二聚体组装方式来检验这个假设 通过 Fe2+ 结合稳定并与端肽基赖氨酸残基相互作用以调节胶原交联形成 和肺部肿瘤的发生。 总之，我的建议将解决肺癌转移的临床问题。新颖之处在于 初步结果提供了对胶原蛋白 LH 的第一个结构见解，挑战了当前的假设 促进 LH 酶活性量化和候选物评估的范式和技术 转移驱动因素。