Acquisition of NMR and X-ray Instrumentation for Structural Biology

结构生物学和生物物理学使用核磁共振光谱学和X射线晶体学是哥伦比亚大学的历史优势。研究兴趣范围从新技术的开发，到蛋白质和核酸的结构和动力学特性的测定，再到生物大分子的基本生物物理和化学特性的基于结构的研究。哥伦比亚大学对结构生物学和生物物理学的开创性贡献的未来延续和扩展受到现有NMR和X射线仪器的年龄增长和不足的威胁。NMR中心目前拥有三台布鲁克NMR光谱仪，质子共振频率分别为300 MHz、400 MHz和500 MHz。光谱仪安装于1985年;只有500 MHz仪器自最初安装以来进行了重大升级。核磁共振波谱仪的年代久远，妨碍了极其重要的现代实验的进行。生物化学和分子生物物理学系的X射线设施包括两台理学RU-200旋转阳极X射线发生器、两台Huber旋进相机、一台理学RAXIS-II成像区域探测器、一台由理学AFC-6测角仪服务的Xuong-Hamlin多线区域探测器和三台低温系统。Xuong-Hamlin多线探测器不再适用于当前的研究项目。对于核磁共振和X射线设施，目前用户安排关键实验的积压时间为一到两个月。 为了满足明显的需求，将购买一台新的600兆赫核磁共振波谱仪，并为500兆赫和400兆赫仪器购买控制台升级。（1）需要额外的高场光谱仪，以便及时进入（新兴）研究小组目前的教师，并使有可能招聘一个或两个额外的初级教师。(2)500 MHz控制台需要升级，以进行最先进的实验。(3)400兆赫光谱仪的升级将使方法的发展，初步数据收集，以评估样品的完整性，并在较低的领域进行培训。这将极大地利用500 MHz和600 MHz频谱仪的生产。 为了减轻目前X射线晶体学仪器的局限性，将购买两个成像板面积探测器系统（包括探测器、聚焦镜和低温输送系统）和一个理学RU-200旋转阳极X射线发生器。根据要求，X射线设施将容纳三个X射线发生器，三个区域探测器系统和两个Huber旋进相机。每个X射线发生器可支持两个旋进相机或区域探测器;因此，其中两个X射线发生器将配备一个旋进相机和一个区域探测器。第三台发电机将配备一个区域探测器，为将来的扩展留出一个插槽。Xuong-Hamlin探测器将退役。所要求的仪器将产生下列优点：（1）X射线设备的能力大约增加一倍，将使现有的研究小组能够及时进行实验;（2）由于有足够的X射线仪器来支持另一个新的研究小组，将有助于在生物分子X射线晶体学方面招聘一名新的教员;以及（3）新探测器（和光学器件）的增加的灵敏度将允许先前在同步加速器源处进行的实验的“内部”性能。

Structural biology and biophysics using NMR spectroscopy and x-ray crystallography are historic strengths of Columbia University. Research interests range from the development of new techniques, to the determination of the structural and dynamical properties of proteins and nucleic acids, to the structure-based investigation of fundamental biophysical and chemical properties of biological macromolecules. Future continuation and expansion of Columbia University's seminal contributions to structural biology and biophysics is threatened by the increasing age and inadequacy of the existing NMR and x-ray instrumentation. The NMR Center presently houses three Bruker NMR spectrometers with proton resonance frequencies of 300 MHz, 400 MHz and 500 MHz. The spectrometers were installed in 1985; only the 500 MHz instrument has been upgraded to any significant extent since the original installation. The age of the NMR spectrometers precludes performance of critically important modern experiments. The Department of Biochemistry and Molecular Biophysics x-ray facility houses two Rigaku RU-200 rotating anode x-ray generators, two Huber precession cameras, one Rigaku RAXIS-II imaging area detector, one Xuong-Hamlin multiwire area detector served by a Rigaku AFC-6 goniormeter, and three cryogenic systems. The Xuong-Hamlin multiwire detector is no longer suitable for current research projects. For both the NMR and x-ray facilities, backlogs for scheduling critical experiments ranges from one to two months with current users. To address evident needs, a new 600 MHz NMR spectrometer will be purchased and consoles upgrades will be purchased for the 500 MHz and 400 M-Hz instruments .These decisions are based on the following considerations: (1) an additional high field spectrometer is necessary to provide timely access for the (burgeoning) research groups of the present faculty and to make possible recruitment of one or two additional junior faculty members. (2) An upgr ade of the 500 MHz console is required for performance of state-of-the-art experiments. (3) An upgrade of the 400 MHz spectrometer will allow methods development, preliminary data collection to evaluate sample integrity, and training to be performed at lower field. This will have the effect of significantly leveraging production on the 500 MHz and 600 MHz spectrometers. To alleviate the limitations of the present x-ray crystallographic instrumentation, two imaging plate area detectors systems (including detector, focusing mirrors and cryo delivery system) and one Rigaku RU-200 rotating anode x-ray generator will be purchased. Following the requested acquisitions, the x-ray facility will house three x-ray generators, three area detector systems, and two Huber precession cameras. Each x-ray generator can support two precession cameras or area detectors; therefore, two of the x-ray generators will be equipped with a precession camera and an area detector. The third generator will be equipped with an area detector, leaving one slot for future expansion. The Xuong-Hamlin detector will be decommissioned. The following advantages will accrue from the requested instrumentation: (I) the approximate doubling of the capacity of the x-ray facility will permit existing research groups to perform experiments in a timely fashion; (2) recruitment of an additional faculty member in biomolecular x-ray crystallography will be facilitated by the availability of sufficient x-ray instrumentation to support another beginning faculty research group; and (3) the increased sensitivity of the new detectors (and optics) will permit performance "in-house" of experiments that previously have been performed at synchrotron sources.