800 MHz NMR SPECTROMETER

描述（由申请人提供）：得克萨斯大学西南医学中心在达拉斯已开始在其核磁共振教师和设施的重大扩张。为了推动这一计划，该大学已承诺超过500万美元用于购买额外的仪器，建设最先进的设施，雇用专门的支持人员和任命高级教师。该计划的组成部分是购买一台配备冷冻探针的800 MHz光谱仪，以支持无法在较低场仪器上进行的新研究计划，并要求提供资金以协助这一奋进。该仪器将支持生物化学和药理学系四个主要NMR组的30个用户的研究，以及大学内外的众多合作者和小用户。使用该仪器的研究将解决广泛的生物学领域的核心问题，包括神经递质释放和组成性细胞内膜交通的分子机制，通过集成信号网络控制肌动蛋白细胞骨架结构和动力学，调节细胞缺氧反应和昼夜节律循环，以及确保有丝分裂中染色体高保真分离的有丝分裂检查点的分子机制。在这些领域的具体结构/生物物理问题包括钙传感器，突触结合蛋白，及其与SNARE蛋白，膜融合机制的基本组成部分的相互作用的研究; 240 kDa Arp 2/3复合物，在细胞中的中央肌动蛋白成核机的激活机制，通过自抑制蛋白的信号整合的生物物理基础; PAS结构域通过天然和非天然配体诱导的蛋白质结构和动力学变化来控制生物学功能的能力;有丝分裂检查点蛋白Mad 2通过其上游调节因子Mad 1及其下游靶点Cdc 20竞争。这些研究将需要对15-250 kDa的多组分系统进行NMR分析，并将涉及CRIPT/TROSY方法的创新使用、镧系元素辅助磁场排列、多场动力学研究和极稀溶液分析，需要最大场强和灵敏度。总之，UT Southwestern的800 MHz光谱仪将促进重要生物领域的前沿科学，将NMR方法的边界扩展到大型和/或复杂系统。该工具将得到联合国大学的大力支持，并通过一项确保有效使用和维护的详细计划、培训新用户的具体机制以及向更广泛的社区传播成果来管理。

DESCRIPTION (provided by applicant): The University of Texas Southwestern Medical Center in Dallas has initiated a major expansion in its NMR faculty and facilities. To drive this program, the University has committed over $5M for purchase of additional instruments, construction of state-of-the-art facilities, hiring of dedicated support personnel and appointment of senior faculty. Integral to this program is the purchase of an 800 MHz spectrometer equipped with a cryoprobe to support new research initiatives that cannot be carried out on lower field instruments, and funds are requested to assist in this endeavor. This instrument will support the research of 30 users in the four principal NMR groups in the Departments of Biochemistry and Pharmacology, plus numerous collaborators and minor users within and outside the university. Research using the instrument will address central questions in a broad range of biological areas, including molecular mechanisms of neurotransmitter release and constitutive intracellular membrane traffic, control of actin cytoskeletal architecture and dynamics through integrated signaling networks, regulation of cellular hypoxia responses and circadian cycling, and molecular mechanisms of a mitotic checkpoint that ensures the high-fidelity segregation of chromosomes in mitosis. Specific structural/biophysical problems in these areas include studies of the calcium sensor, synaptotagmin, and its interaction with SNARE proteins, essential components of the membrane fusion machinery; the activation mechanism of the 240 kDa Arp2/3 complex, the central actin nucleating machine in the cell, the biophysical foundations of signal integration by autoinhibitory proteins; the ability of PAS domains to control biological function through changes in protein structure and dynamics induced by natural and non-natural ligands; competition for the mitotic checkpoint protein Mad2 by its upstream regulator, Mad1, and its downstream target, Cdc20. These studies will require NMR analyses of multi-component systems ranging in size from 15-250 kDa, and will involve innovative use of CRIPT/TROSY methods, lanthanide-assisted magnetic field alignment, multi-field dynamics studies and analysis of very dilute solutions, necessitating maximum field strength and sensitivity. In summary, an 800 MHz spectrometer at UT Southwestern will foster cutting edge science in important biological areas that will expand the boundaries of NMR methods to large and/or complex systems. The instrument will be strongly supported by the University, and administered through a detailed plan that ensures efficient use and maintenance, specific mechanisms for training new users and dissemination of results to the larger communitv.