ANALYTICAL ULTRACENTRIFUGE FOR MACROMOLECULAR STUDIES

用于大分子研究的分析超速离心机

• 批准号: 6292022
• 负责人: Cynthia Vianne Stauffacher
• 金额: $ 27.06万
• 依托单位: PURDUE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-03-01 至 2002-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6292022
• 关键词: analytical ultracentrifugation; biomedical equipment purchase; intermolecular interaction; macromolecule

The properties and capabilities are described of an analytical ultracentrifuge, Beckman Optima XL-I, with both interference and absorption optics, that will be used by a group of Purdue research scientists to investigate the interactions between macromolecules in biologically important systems. The acquisition of this instrument is part of a plan to assemble an array of instruments in a center for biophysical studies of macromolecules. This array of instruments currently includes a surface plasmon resonance instrument, an isothermal calorimeter, and a ten-year old Beckman Model XL-A analytical ultracentrifuge. Future development of this center targets the addition of state-of-the-art fluorescence, FTIR and Raman spectrometers, plus a stopped flow rapid reaction instrument. This proposal for the XL-I analytical ultracentrifuge is submitted by a group of six investigators involved in studies of protein-protein, protein-nucleic acid, protein-ligand and protein- membrane interactions. These investigators have expertise in high resolution crystallographic structural studies as well as in biochemistry, biophysics and molecular genetics, and are interested in complementing their structural results with the biophysical measurements that can illuminate the details of assembly of large complexes, changes in interaction states with ligand substrates or effectors or with modifications that mimic cellular signaling states. Investigations of stability of large complexes and search for conditions of monodisperse preparations of membrane proteins are also proposed, as an aid to the crystallization of these difficult systems. Initial studies with the Beckman XL-A have demonstrated the usefulness of analytical ultracentrifugation in a wide variety of systems including: the binding affinity of membrane proteins for their soluble counterparts, assembly of animal viruses, signal transduction protein interactions, interactions between toxins and their cellular receptors as well as the assembly of large protein complexes for transmembrane transport. The XL-I will extend the range of studies into those involving weaker interactions as well as to studies where the optical characteristics of the samples are inappropriate for the absorption optics of the XL-A and will compliment and extend the information obtained by both structural and biophysical methods at Purdue.

介绍了贝克曼Optima XL-I分析型超离心机的特性和性能，该离心机具有干涉和吸收光学，将被普渡大学的一组研究人员用于研究生物重要系统中大分子之间的相互作用。这台仪器的收购是在大分子生物物理研究中心组装一系列仪器计划的一部分。该阵列的仪器目前包括一个表面等离子体共振仪，一个等温量热计，和一个十年的贝克曼模型XL-A分析超离心机。该中心未来的发展目标是增加最先进的荧光，FTIR和拉曼光谱仪，以及一个停流快速反应仪器。这个关于XL-I分析型超离心机的建议是由参与蛋白质-蛋白质、蛋白质-核酸、蛋白质-配体和蛋白质-膜相互作用研究的六名研究人员提交的。这些研究人员在高分辨率晶体结构研究以及生物化学、生物物理学和分子遗传学方面具有专业知识，并且对用生物物理测量来补充他们的结构结果感兴趣，这些测量可以阐明大型复合物组装的细节，与配体底物或效应器相互作用状态的变化，或与模拟细胞信号传导状态的修饰。研究大型配合物的稳定性和寻找膜蛋白单分散制备的条件也被提出，以帮助这些困难的系统结晶。贝克曼XL-A的初步研究已经证明了分析超离心在各种系统中的有用性，包括：膜蛋白与可溶性对应物的结合亲和力，动物病毒的组装，信号转导蛋白的相互作用，毒素与其细胞受体之间的相互作用，以及跨膜运输的大蛋白复合物的组装。XL-I将把研究范围扩展到那些涉及弱相互作用的研究，以及那些样品的光学特性与XL-A的吸收光学不合适的研究，并将补充和扩展普渡大学通过结构和生物物理方法获得的信息。