MADMAX: PRECISE MEASUREMENT OF CONFORMATIONAL CHANGES IN PROTEINS

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 该项目涉及开发一种新的方法，即使用中角X射线溶液散射的多波长反常衍射(MADMAX)，用于精确测量蛋白质和其他水溶液中的大分子之间的原子间距离。它将使现有技术无法研究的结构变化和分子内运动的表征成为可能。蛋白质是动态的分子，其活动对所有生物过程都有贡献。几乎所有的蛋白质功能都需要分子内运动，无论是施加机械力、化学转化还是分子移位。MadMax将有可能对溶液中蛋白质的这些运动进行精确的原子水平测量。MADMAX应该普遍适用于溶液中的大分子，并能够在包括变构相互作用、蛋白质-配体相互作用、通道门控、结构域移动和蛋白质折叠在内的广泛现象中阐明蛋白质的作用。它应该能够测量原子间矢量长度的变化，精度高达~1°。它可以被改装成至少毫秒级的时间分辨率研究。 该项目的目标是： (I)演示如何准确测量具有良好特性的蛋白质在溶液散射中的异常差异。 (Ii)从原子坐标集预测异常差异。 (3)确定该方法的局限性：该方法适用于多大的蛋白质？检测需要多少个标签？标签位置的混乱程度可以容忍多少？ (Iv)使用MadMax精确测量由于配体结合而引起的结构变化。 (V)利用MadMax研究膜蛋白的结构变化。 该项目涉及开发一种新的方法，即使用中角X射线溶液散射的多波长反常衍射(MADMAX)，用于精确测量蛋白质和其他水溶液中的大分子之间的原子间距离。它将使现有技术无法研究的结构变化和分子内运动的表征成为可能。蛋白质是动态的分子，其活动对所有生物过程都有贡献。几乎所有的蛋白质功能都需要分子内运动，无论是施加机械力、化学转化还是分子移位。MadMax将有可能对溶液中蛋白质的这些运动进行精确的原子水平测量。MADMAX应该普遍适用于溶液中的大分子，并能够在包括变构相互作用、蛋白质-配体相互作用、通道门控、结构域移动和蛋白质折叠在内的广泛现象中阐明蛋白质的作用。它应该能够测量原子间矢量长度的变化，精度高达~1°。它可以被改装成至少毫秒级的时间分辨率研究。 该项目的目标是： (I)演示如何准确测量具有良好特性的蛋白质在溶液散射中的异常差异。 (Ii)从原子坐标集预测异常差异。 (3)确定该方法的局限性：该方法适用于多大的蛋白质？检测需要多少个标签？标签位置的混乱程度可以容忍多少？ (Iv)使用MadMax精确测量由于配体结合而引起的结构变化。 (V)利用MadMax研究膜蛋白的结构变化。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. This project involves development of a new method, Multi-wavelength Anomalous Diffraction using Medium Angle X-ray solution scattering (MADMAX), for the precise measurement of interatomic distances within proteins and other macromolecules in aqueous solution. It will make possible the characterization of structural changes and intra-molecular movements that cannot be studied by existing techniques. Proteins are dynamic molecules whose activities contribute to all biological processes. Virtually all protein function requires intra-molecular movement, whether for the application of mechanical force, chemical transformation or molecular translocation. MADMAX will make possible accurate atomic-level measurement of these movements within proteins in solution. MADMAX should be generally applicable to macromolecules in solution and capable of elucidating protein action during a broad range of phenomena including allosteric interactions; protein-ligand interactions; channel gating; domain movements; and protein folding. It should be capable of measuring changes in the length of interatomic vectors with an accuracy of up to ~1 ¿. It can be adapted for time-resolved studies  to at least millisecond resolution. The goals of this project are: (i) Demonstrate the accurate measurement of anomalous differences in solution scattering from well characterized proteins. (ii) Predict the anomalous differences from atomic coordinate sets. (iii) Determine the limitations of the method: How large a protein is this method applicable to? How many labels are required for detection? How much disorder in the label position can be tolerated? (iv) Use MADMAX for precise measurement of structural changes due to ligand binding. (v) Use MADMAX for the study of structural changes in membrane proteins. This project involves development of a new method, Multi-wavelength Anomalous Diffraction using Medium Angle X-ray solution scattering (MADMAX), for the precise measurement of interatomic distances within proteins and other macromolecules in aqueous solution. It will make possible the characterization of structural changes and intra-molecular movements that cannot be studied by existing techniques. Proteins are dynamic molecules whose activities contribute to all biological processes. Virtually all protein function requires intra-molecular movement, whether for the application of mechanical force, chemical transformation or molecular translocation. MADMAX will make possible accurate atomic-level measurement of these movements within proteins in solution. MADMAX should be generally applicable to macromolecules in solution and capable of elucidating protein action during a broad range of phenomena including allosteric interactions; protein-ligand interactions; channel gating; domain movements; and protein folding. It should be capable of measuring changes in the length of interatomic vectors with an accuracy of up to ~1 ¿. It can be adapted for time-resolved studies  to at least millisecond resolution. The goals of this project are: (i) Demonstrate the accurate measurement of anomalous differences in solution scattering from well characterized proteins. (ii) Predict the anomalous differences from atomic coordinate sets. (iii) Determine the limitations of the method: How large a protein is this method applicable to? How many labels are required for detection? How much disorder in the label position can be tolerated? (iv) Use MADMAX for precise measurement of structural changes due to ligand binding. (v) Use MADMAX for the study of structural changes in membrane proteins.