800 MHZ SPECTROMETER FOR BIOMOLECULAR NMR: CANCER

这个子项目是利用由NIH/NCRR资助的共享仪器赠款提供的资源的许多研究子项目之一。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是用于拨款的，而不一定是用于调查人员的机构。描述(由申请人提供)：来自密苏里大学哥伦比亚、圣路易斯和堪萨斯城校区以及华盛顿大学圣彼得堡分校的七组核磁共振用户。路易斯为密苏里州寻找第一台800兆赫核磁共振波谱仪。高场光谱仪将在他们的NIH项目中引入更重要的结构生物学。密苏里-哥伦比亚大学(UMC)位于从圣路易斯到堪萨斯城的生命科学走廊的地理中心，将托管800 MHz系统。决定800人政策的咨询委员会将有从圣路易斯到堪萨斯城的广泛代表。统一适用于所有用户的收费制度将收回800名用户的大部分运营成本。联华电子提供了一个大型配套套餐和新的空间，专门为在其医学院的新楼内举办800人而量身定做。申请的资金将用于一台带有18.8T屏蔽磁铁的800兆赫光谱仪。具有突出灵敏度的低温探头仍然是一个额外的高度优先事项。所有这些现在使用600 MHz设备的团队都需要800 MHz带来的灵敏度、分辨率和TROSY线锐化方面的提升。四个组织将雇用这800人从事与癌症相关的工作。Van Doren小组将研究(I)金属蛋白酶与抑制物和底物蛋白的相互作用，以及(Ii)转化的肿瘤病毒蛋白与宿主细胞的目标蛋白的相互作用。Ma和Deutscher将确定Galectin-3与肿瘤靶向肽和全长Galectin-3二聚体结合的核磁共振结构。Ellen Li的团队将研究配体如何改变维甲酸X受体(RXR)配体结合域的平衡动力学。约翰-斯蒂芬·泰勒的团队将对皮肤癌背后的DNA照相产品获得更详细的结构洞察。亨兹尔将研究内耳听觉器官中主要蛋白质(OCP2和OCP1)的结构特征和相互作用，内耳听觉器官是一种结合连接蛋白26的复合体，在遗传性耳聋中最常发生突变。三个小组将研究DNA结合蛋白组装。Dupureur的团队将对限制性内切酶的动力学和构象调整进行第一次核磁共振研究，以响应DNA结合和“STAR活性”的条件。莱伊特将研究金属反应转录因子-1的锌感应DNA结合域的结构和动力学，它调节哺乳动物的锌稳态。他将研究它的所有六个自由的和与DNA绑定的锌手指。Dreyfus和Laity将描述来自致病细菌的细胞毒性镉B毒素的结构和动力学，这些毒素是游离的和与DNA结合的。在哺乳动物细胞中，CDtB亚基足以引起DNA损伤和细胞周期停滞。

This subproject is one of many research subprojects utilizing the resources provided by a Shared Instrumentation 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 grant, which is not necessarily the institution for the investigator. DESCRIPTION (provided by applicant): Seven groups of NMR users from the Columbia, St. Louis, and Kansas City campuses of the University of Missouri and from Washington University-St. Louis seek the first 800 MHz NMR spectrometer for the state of Missouri. The high field spectrometer will introduce structural biology of greater significance to their NIH projects. The University of Missouri-Columbia (UMC), in the geographical center of the life sciences corridor from St. Louis through Kansas City, will host the 800 MHz system. The advisory committee for deciding policy for the 800 will have broad representation from St. Louis to Kansas City. A fee system, uniformly applied among all users, will recover most of the operating costs of the 800. UMC offers a large matching package and new space tailored to hosting the 800 within a new building of its School of Medicine. The funds requested will be applied to an 800 MHz spectrometer with a shielded 18.8 T magnet. A cryogenic probe for outstanding sensitivity remains an additional high priority. The boosts in sensitivity, resolution, and TROSY line sharpening afforded by 800 MHz are needed by all these groups now using 600 MHz equipment. Four groups will employ the 800 for cancer-related work. The Van Doren group will investigate (i) interactions of metalloproteases with inhibitor and substrate proteins and (ii) interactions of transforming tumor viral proteins with their target proteins of host cells. Ma and Deutscher will determine NMR structures of galectin-3 bound to a tumor-targeting peptide and a full-length galectin-3 dimer. Ellen Li's group will investigate how ligands shift the equilibrium dynamics of retinoid-X-receptor (RXR) ligand binding domain. John-Stephen Taylor's group will obtain more detailed structural insights on DNA photoproducts underlying skin cancer. Henzl will study structural features and interactions of principal proteins (OCP2 and OCP1) in the auditory organ of the inner ear, a complex that binds connexin 26 most often mutated in hereditary deafness. Three groups will study DNA-binding protein assemblies. Dupureur's group will undertake the first NMR investigations of dynamics and conformational adjustments of a restriction endonuclease in response to DNA binding and conditions of "star activity". Laity will study structure and dynamics of the zinc-sensing DNA-binding domain of metal-responsive transcription factor-1 that regulates zinc homeostasis in mammals. He will study all six of its zinc fingers free and bound to DNA. Dreyfus and Laity will characterize the structure and dynamics of cytolethal CdtB toxin from pathogenic bacteria, free and bound to DNA. The CdtB subunit is sufficient to cause DNA damage and to arrest the cell cycle in mammalian cells.