Proj 1: Improving macromolecule crystallization using symmetry-inducing GFP

项目 1：使用对称性诱导 GFP 改善大分子结晶

• 批准号: 8471726
• 负责人: GEOFFREY S. WALDO
• 金额: $ 50.94万
• 依托单位: LOS ALAMOS NAT SECTY-LOS ALAMOS NAT LAB
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8471726
• 关键词: Binding; Crystallization; Engineering; Green Fluorescent Proteins; Link; Methods; Mutation; Probability; Proteins; Series; Structure; Testing; Time; Variant; X-Ray Crystallography; abstracting; base; enhanced green fluorescent protein; improved; link protein; macromolecule; protein complex; reconstitution

Project 1. Summary/Abstract Improving macromolecular crystallization using lattice-promoting variants of GFP The bottleneck in structure determination by X-ray crystallography is crystallization, where -70% of purified proteins fail. A major reason proteins fail to crystallize is insufficient suitable lattice-forming contacts. The central hypothesis in this project is that the probability of obtaining crystals is increased every time a new form of a molecule with new potential crystal contacts is tested. We will develop rapid and general methods to generate numerous venations on a protein molecule by linking the protein to a series of oligomeric forms of green fluorescent protein (GFP). The use of GFP is a major advantage because it can be linked firmly to practically any protein by inserting a hairpin (two strands connected by a loop) from GFP into a loop of the target protein. The target protein containing this extra hairpin can bind tightly to a

项目1.摘要/摘要 利用GFP晶格促进变异体改善大分子结晶 在X射线结晶学结构测定中的瓶颈是结晶，其中-70%的纯化蛋白质失效。蛋白质不能结晶的一个主要原因是没有合适的晶格形成接触。这个项目的中心假设是，每当测试一种具有新的潜在晶体接触的新形式的分子时，获得晶体的可能性就会增加。我们将开发快速和通用的方法，通过将蛋白质与一系列低聚形式的绿色荧光蛋白(GFP)联系起来，在蛋白质分子上产生大量脉络。使用GFP是一个主要的优势，因为它可以通过将发夹(由环连接的两条链)插入到目标蛋白质的环中而牢固地连接到几乎任何蛋白质上。含有这种额外发夹的目标蛋白可以紧密结合到