Anomalous Diffraction Analysis of Biomolecular Structure

生物分子结构的反常衍射分析

基本信息

批准号：
8697562
负责人：
WAYNE A. HENDRICKSON
金额：
$ 46.37万
依托单位：
COLUMBIA UNIVERSITY HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-06-03 至 2018-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8697562
关键词：
Address Automation Basic Science Biological Calcium Caring Cluster Analysis Communities Complex Computer software DNA Data Data Collection Data Set Development Drug Formulations Electrons Evaluation Health Heart failure Human Indium Infection Lasers Light Malignant Neoplasms Measures Medical Membrane Proteins Methods Modeling Molecular Chaperones Neurodegenerative Disorders New York Noise Phase Philosophy Phosphorus Probability Procedures Process Proteins Radiation Refractory Research Resolution Signal Transduction Solutions Solvents Source Structure Sulfur Synchrotrons System Testing absorption base beamline biological systems computerized tools design detector experience improved instrumentation macromolecule meetings method development microbial novel strategies programs protein-histidine kinase public health relevance radiation effect receptor research study structural biology structural genomics technology development three dimensional structure tool

项目摘要

DESCRIPTION (provided by applicant): The overall objective of the proposed research is to develop enhanced anomalous diffraction methods for analyzing structures of biological macromolecules. We build on our recent accomplishments at New York Structural Biology Center (NYSBC) beamlines at Brookhaven's National Synchrotron Light Source (NSLS) and worldwide experience showing that multiwavelength anomalous diffraction (MAD) and SAD, MAD's single-wavelength counterpart, now predominate for de novo determinations of three- dimensional structures for biological macromolecules. We propose to optimize SAD and MAD phasing procedures by meeting the demands of compelling applications to current problems of biological significance. Biologically exciting problems motivate the development of appropriate tools, and forefront methods accelerate the solution of structures for systems of biological and medical significance. The overall objective is embodied in four specific aims: (1) we propose to enhance SAD phasing procedures for challenging problems such as selenomethionyl proteins at low resolution and only-light-atom native structures. A focus is on improved methods for increasing signal-to-noise ratios for anomalous diffraction by combining data from many crystals. (2) We propose to enhance MAD procedures for accurate experimental phase evaluation. Data collection strategies will be devised to mitigate radiation damage and minimize systematic errors. (3) We propose to develop procedures for low-energy anomalous diffraction experiments. We will develop experimental procedures and design instrumentation for low-energy (3 - 7 keV) experiments aiming to enhance anomalous signal from light atoms such as sulfur and phosphorous. (4) We propose to develop automation and enhanced procedures to facilitate SAD and MAD analyses from many crystals. A first priority is to encode and disseminate our current multi-crystal SAD process in convenient software.

描述（由申请人提供）：所提议研究的总体目标是开发用于分析生物大分子结构的增强型反常衍射方法。我们以布鲁克海文国家同步加速器光源 (NSLS) 的纽约结构生物学中心 (NYSBC) 光束线最近取得的成就为基础，以及全球经验表明，多波长反常衍射 (MAD) 和 SAD（MAD 的单波长对应物）现在在 de novo 领域占主导地位生物大分子三维结构的测定。我们建议通过满足当前具有生物学意义的问题的引人注目的应用需求来优化 SAD 和 MAD 定相程序。生物学上令人兴奋的问题激发了适当工具的开发，前沿方法加速了具有生物学和医学意义的系统结构的解决。总体目标体现在四个具体目标中：（1）我们建议增强 SAD 定相程序，以解决低分辨率的硒代甲硫氨酰蛋白和仅轻原子天然结构等具有挑战性的问题。重点是通过组合来自许多晶体的数据来提高反常衍射的信噪比的改进方法。 (2) 我们建议加强 MAD 程序以实现准确的实验阶段评估。将制定数据收集策略以减轻辐射损害并最大限度地减少系统误差。 (3) 我们建议开发低能反常衍射实验程序。我们将开发低能（3 - 7 keV）实验的实验程序和设计仪器，旨在增强硫和磷等轻原子的异常信号。 (4) 我们建议开发自动化和增强程序，以促进许多晶体的 SAD 和 MAD 分析。首要任务是在方便的软件中编码和传播我们当前的多晶 SAD 过程。