Characterization and Crystallization of Pathogenic Protozoa Proteins

致病性原生动物蛋白的表征和结晶

• 批准号: 7071314
• 负责人: WILHELMUS G. J. HOL
• 金额: $ 32.32万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-01 至 2009-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7071314
• 关键词: X ray crystallography; antiprotozoal agents; bioimaging /biomedical imaging; bioinformatics; chemical structure function; crystallization; drug discovery /isolation; high throughput technology; information systems; iodination; ligands; light scattering; molecular site; nucleic acid methylation; peptide library; protein structure function; proteolysis; proteomics; robotics; structural biology

The overall goal of the Medical Structural Genomics of Pathogenic Protozoa (MSGPP) Program Project is to determine crystal structures of biomedically important proteins of pathogenic protozoa in complex with small-molecule ligands, and to use that information for accelerated drug development for treatment and prevention of the diseases caused by the ten major pathogenic organisms targeted. MSGPP is on the crossroads of the genomes sequencing and structural genomics revolutions in translating nucleotide sequence data into proteins, crystals, three-dimensional structures and protein-inhibiting compounds. This Project 3 is critical for this tightly integrated Program Project since it is at the intersection of Project 1, which will provide Ligands, and Projects, which will provide numerous MSGPP protein samples, while the major output of this Project will be crystals if protein-ligand complexes for X-ray structure determination in Project 4. We will use multiple avenues to obtain X-ray diffraction quality crystals. In a first step, several characterization steps will be performed to see how the protein can be trimmed, stabilized with ligands, provided with a low-polydispersity inducing buffer, and chemically modified. In a second step, a series of complementary approaches and in particular an integrated set of robots and databases will be applied to grow crystals in conjunction with several focused optimization methodologies. Innovative collaborations are in place for high-throughput initial crystal screening, determination of second virial coefficients, capillary crystallization robotics, and automated crystal image analysis. The entire spectrum of approaches within MSGPP is estimated to result in an overall crystallization success rate between 30 to 40% for each group of target variants.

致病原生动物（MSGPP）计划项目的医学结构基因组学项目的总体目标是确定与小分子配体复合物中生物医学上重要的致病原生动物蛋白质的晶体结构，并将这些信息用于加速药物开发和预防药物开发和预防。由十种主要的病原生物靶向的疾病引起的疾病。 MSGPP在将核苷酸序列数据转化为蛋白质，晶体，三维结构和抑制蛋白质的化合物的基因组测序和结构基因组学的十字架上。该项目3对于这个紧密整合的程序项目至关重要，因为它是在项目1的交集中，该项目将提供配体和项目，该项目将提供许多MSGPP蛋白质样品，而该项目的主要输出将是晶体，如果蛋白质 - 项目4中的X射线结构确定的配体配合物。我们将使用多个途径获得X射线衍射质量晶体。在第一步中，将执行几个表征步骤，以查看如何用配体稳定蛋白质，并提供诱导缓冲液的​​低多分散性和化学修饰的蛋白质。在第二步中，将采用一系列补充方法，特别是一组集成的机器人和数据库集与多种集中优化方法结合起来，将晶体延长。进行高通量初始晶体筛选，第二个病毒系数，毛细管的确定，进行了创新的合作 结晶机器人和自动晶体图像分析。估计MSGPP中的整个方法估计会导致整体结晶成功 每组目标变体的率在30至40％之间。