ANALYTICAL ULTRACENTRIFUGE FACILITY

共享分析超速离心设备的建立将加强和扩大NIH支持的UMDNJ-Robert Wood Johnson医学院的一些研究人员的研究计划。其中一些研究人员以生物物理学为导向，而另一些以分子生物学为重点的研究人员在我们科学界强大的结构生物学实验室的支持下，越来越多地用结构表征来补充他们的功能研究。本申请中提出的沉降研究将提供从模型三螺旋和盘状多肽到调节蛋白、转录和延伸因子复合体等系统中的分子质量、形状、化学计量和分子间相互作用的信息。这些研究人员对将沉积方法应用于他们的研究项目的兴趣受到两个因素的推动。首先，最近分析性超离心机设计和光学技术的进步，以及数据采集和分析的自动化，使得有积极性的科学家以及敬业的专家都能使用这种方法。其次，由于高效的表达系统、固态多肽合成和改进的纯化方法，用于提高灵敏度的分析性超速离心所需的纯化分子和络合物的数量越来越多。拟议的沉淀法研究结果具有严格的热力学基础，将提供补充现有方法的信息，并可与核磁共振光谱和X射线结晶学获得的分子结构数据相结合。此外，它是研究超高相对分子质量络合物和其他方法结果不明确的系统的首选技术。这些研究人员对分析超速离心机的需求在现有条件下是无法满足的。在新泽西州，没有一家学术机构拥有这样的仪器，合作只能提供有限的沉淀物研究机会。关于这一应用的几名调查人员参与了与国家设施或私人实验室的超速离心机专家的富有成效的合作。但这种合作因其远距离性质、每个中心正在进行的众多项目以及探索新领域或跟进有趣观察或问题的障碍而受到阻碍。这个应用程序上的任何用户都不需要超过15%-20%的时间使用分析超速离心机，因此任何一个人购买它都是不合理的，然而这种仪器的存在将极大地加强所有用户的研究计划。共享的分析超速离心设备将通过允许罗伯特·伍德·约翰逊医学院的研究人员利用这一强大的技术来加强他们的研究项目。

The establishment of a shared analytical ultracentrifuge facility would enhance and expand the research programs of a number of NIH supported researchers at UMDNJ-Robert Wood Johnson Medical School. Some of these researchers are biophysically oriented, while others with a molecular biology focus have increasingly complemented their functional studies with structural characterization, supported by the strong structural biology laboratories in our scientific community. The sedimentation studies proposed in this application would provide information on molecular mass, shape, stoichiometry, and intermolecular interactions in systems ranging from model triple-helix and coiled-coil peptides to regulatory proteins, transcription and elongation factor complexes. The interest of these investigators in applying sedimentation methods to their research projects has been driven by two factors. Firstly, the recent advances in analytical ultracentrifuge design and optics, as well as the automation of data acquisition and analysis, has made the method accessible to motivated scientists as well as dedicated experts. Secondly, as a result of efficient expression systems, solid state peptide synthesis, and improved purification methodologies, the quantities of purified molecules and complexes necessary for analytical ultracentrifuges with improved sensitivities have increasingly become available. The results of the proposed studies using sedimentation, with its rigorous thermodynamic basis, will provide information complementary to existing methodologies and can be integrated with the molecular structure data obtained from NMR spectroscopy and x-ray crystallography. In addition, it is the technique of choice for studies on very high molecular weight complexes and for systems where other methods have yielded ambiguous results. The need of these researchers for an analytical ultracentrifuge cannot be met under existing conditions. There is no academic institution in New Jersey that has such instrumentation and collaborations have provided only limited access to sedimentation studies. Several investigators on this application have been involved in productive collaborations with ultracentrifuge experts at national facilities or in private laboratories. But such collaborations are hampered by their long distance nature, the numerous projects ongoing at each center, and the barriers to exploring new areas or following up interesting observations or problems. No user on this application requires more than 15-20 percent of the time on an analytical ultracentrifuge, so that its purchase by any one individual cannot be justified, and yet the presence of this instrument would greatly strengthen the research programs of all users. A shared analytical ultracentrifugation facility would strengthen the research projects of Robert Wood Johnson Medical School researchers by allowing them to utilize this powerful technique.