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: 8268582
• 负责人: ANDREW BRADBURY
• 金额: $ 119.65万
• 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/8268582
• 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. PUBLIC HEALTH RELEVANCE: 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. This new structural information 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，其中大约70％的纯化蛋白质失败。蛋白质未能进入结晶状态的两个主要原因是晶格接触太少，并且具有多构象，这通常是丢失的伴侣蛋白的结果。该项目的目的是克服结晶的障碍。该项目中的关键总体想法是，通过（1）以高度适合结晶的形式呈现这种大分子的结晶可以改善这种大分子，并且（2）创建许多不同形式的大分子。这些优化结晶性和变化的主题对于该项目的所有三个组成部分都是至关重要的。我们建议开发的创新新方法将结合自然结合伙伴和结合模块的组合，通过（​​1）创建许多不同形式的分子来改善目标大分子的结晶，以稳定和增强目标大分子的结晶能力，以结晶型分子和（2）找到天然伴侣的晶体。将使用一系列可以与靶分子链接的对称形成模块来创建许多形式的分子。这些模块将基于抗体或绿色荧光蛋白。新型生物信息学和实验方法的结合将发现自然结合伴侣。转化方法是可扩展的，协同的，并通过提供一个分子工具包来多样化靶向蛋白的潜在结晶排列和对称性，从而在结构生物学和结构基因组学上均可加速产生结构生物学。该计划项目将由我们的UCLA/ LOS ALAMOS团队作为三个紧密整合的副标题进行，每个项目都涉及来自UCLA和Los Alamos的研究人员。这项工作将导致结构生物学界将使用的方法来确定蛋白质的结构，从而增加我们对人类健康的理解和治愈人类疾病的能力。 公共卫生相关性：这项工作将导致结构生物学界将使用的方法来确定蛋白质的结构，从而增加我们对人类健康的理解和治愈人类疾病的能力。这种新的结构信息将增加我们对人类健康的理解和治愈人类疾病的能力。