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: 10261533
• 负责人: Houfu Guo
• 金额: $ 24.47万
• 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/10261533
• 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; bone; career development; clinical application; crosslink; design; dimer; human model; inhibitor/antagonist; 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.

项目概要/摘要： 候选人：郭博士在生物化学和结构生物学方面接受过广泛的培训。他是博士后 MD安德森癌症中心的研究员，试图了解胶原赖氨酰羟化酶（LH）的结构 特征调节肺癌转移。郭博士是一位高产的研究者，有18位同行评审 论文（其中6篇为第一作者或共同第一作者）。为了表彰他的成就，他获得了许多 在他的博士和博士后培训期间获得的奖项。 职业发展/培训：郭博士在MD安德森的导师包括Jonathan Kurie博士，一位内科医生， 在肺癌生物学和人类肺癌小鼠模型方面具有专长的科学家，以及John Tainer博士， 一位著名的蛋白质晶体学家，研究大分子组装，为药物设计提供信息。此外，本发明的目的是， 博士Guo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， 是最早阐明胶原蛋白如何被LH修饰的人之一。这些调查人员设计了一个训练 该计划以关键科学学科为中心，将增强郭博士在各自领域的能力 这些国家将继续在各个领域开展工作，并提供顺利过渡到独立所需的技能。 研究：我们的研究小组已经表明，LH 2的高表达驱动肺癌转移，并诱导肿瘤细胞凋亡。 肿瘤间质中的胶原交联开关。LH 2是感兴趣的治疗靶点，但选择性LH抑制剂 是不可用的，部分由于缺乏对LH 2的结构洞察的缺陷。我提议的目的 旨在阐明LH 2促进肺癌转移的结构特征。在初步结果中，我 描述了生产人LH 2蛋白的新方法，并测定了其活性和催化剂的结构。 结构域的病毒LH同源物，这揭示了一个同源二聚体稳定的Fe+2结合。我发现， 活性位点位于二聚体界面上的深表面裂缝的侧面，表明二聚化产生了胶原蛋白， 结合位点，并且邻近活性位点的碱性残基被定位成与 胶原蛋白上的端肽基酸性残基。根据这些发现，我假设LH 2具有端肽-LH 由于其独特的特征，使其能够形成Fe 2+稳定的二聚结构，与Fe 2+相互作用，从而具有活性 端肽基赖氨酸对胶原蛋白的作用。我将通过确定LH二聚体的组装来检验这一假设。 通过Fe 2+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 和肺肿瘤发生。 总之，我的建议将解决肺癌转移的临床问题。新奇在于 初步结果提供了对胶原蛋白LH的第一个结构见解，挑战了目前的假设， 促进LH酶活性定量和候选药物评价的范例和技术 转移驱动因素。