Enhancing crystallization with binding partners, symmetry and diversity

通过结合伙伴、对称性和多样性增强结晶

• 批准号: 8471721
• 负责人: ANDREW BRADBURY
• 金额: $ 115.46万
• 依托单位: LOS ALAMOS NAT SECTY-LOS ALAMOS NAT LAB
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-06-01 至 2016-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8471721
• 关键词: Affinity; Antibodies; Back; Bacteriophages; Binding; Bioinformatics; Communities; Complex; Crystal Formation; Crystallization; Crystallography; DHFR gene; Dimerization; Diversity Library; Engineering; Failure; Fluorescence; Genomics; Green Fluorescent Proteins; Health; Human; Laboratories; Lead; Libraries; Link; Membrane; Methods; Modeling; Molecular; Molecular Chaperones; Molecular Conformation; Mutation; Procedures; Process; Protein Fragment; Proteins; Protocols documentation; Reagent; Recombinants; Research Personnel; Resolution; Site; Staging; Structure; Surface; Testing; Validation; Variant; Work; X-Ray Crystallography; base; human disease; improved; innovation; macromolecule; novel; programs; protein complex; protein folding; protein structure; reconstitution; scaffold; structural biology; structural genomics

DESCRIPTION (provided by applicant): The bottleneck in structure determination by X-ray crystallography is crystallization, where roughly 70% of purified proteins fail. Two major reasons proteins fail to enter the crystalline state are having too few lattice contacts, and having multipe conformations, often the result of missing partner proteins. The purpose of this project is to overcome barriers to crystallization. The key overall ideas in this project are that crystallizatio of a macromolecule or complex can be improved by (1) presenting this macromolecule in a form that is highly suitable for crystallization and (2) creating many different forms of the macromolecule. These themes of optimizing crystallizability and variation are central to all three components of this projects. The innovation new methods we propose to develop will use a combination of natural binding partners and binding modules to improve crystallization of a target macromolecule by (1) creating many different forms of a molecule to crystallize and (2) finding a natural partner macromolecule that stabilizes and enhances the crystallizability of a target macromolecule. Many forms of a molecule will be created using a panel of symmetry-forming modules that can be linked to the target molecule. These modules will be antibody- or green fluorescent protein-based. Natural binding partners will be found with a combination of novel bioinformatics and experimental approaches. The transformative methods are scalable, synergistic, and offer the potential of accelerating results in both structural biology and structual genomics by providing a molecular toolkit for diversifying the potential crystallization arrangements and symmetries of targeted proteins. This program project will be carried out by our UCLA/ Los Alamos team as three tightly integrated subprojects, each involving researchers from both UCLA and Los Alamos. This work will lead to methods that will be used by the structural biology community to determine structures of proteins that will increase our understanding of human health and our ability to cure human disease.

描述（由申请人提供）：X 射线晶体学确定结构的瓶颈是结晶，大约 70% 的纯化蛋白质在结晶过程中失败。蛋白质无法进入结晶状态的两个主要原因是晶格接触太少，以及具有多种构象，这通常是缺少伴侣蛋白质的结果。该项目的目的是克服结晶障碍。该项目的关键总体想法是，可以通过以下方式改进大分子或复合物的结晶：（1）以高度适合结晶的形式呈现该大分子；以及（2）创建该大分子的许多不同形式。这些优化结晶性和变化的主题是该项目所有三个组成部分的核心。我们建议开发的创新方法将使用天然结合伴侣和结合模块的组合来改善目标大分子的结晶，方法是：（1）创建许多不同形式的分子进行结晶；（2）找到稳定和增强目标大分子结晶性的天然伴侣大分子。许多形式的分子将使用一组可连接到目标分子的对称形成模块来创建。这些模块将基于抗体或绿色荧光蛋白。结合新颖的生物信息学和实验方法将发现天然结合伴侣。这些变革性方法具有可扩展性、协同性，并且通过提供使目标蛋白质的潜在结晶排列和对称性多样化的分子工具包，有可能加速结构生物学和结构基因组学的结果。该计划项目将由我们的加州大学洛杉矶分校/洛斯阿拉莫斯团队作为三个紧密结合的子项目来实施，每个子项目都涉及加州大学洛杉矶分校和洛斯阿拉莫斯的研究人员。这项工作将带来结构生物学界用来确定蛋白质结构的方法，从而增加我们对人类健康的理解和治疗人类疾病的能力。