XL-A Analytical Ultracentrifuge for the Analysis of Protein-Protein Interactions

这是一项收购贝克曼仪器Optima系列XL-A分析超离心机的建议，该离心机配备了紫外线扫描仪和瑞利光学系统。该仪器将取代波士顿生物医学研究所（BBRI）分析超离心研究实验室（AURL）中2台大量使用的Beckman Model-E。其中一架Model-E目前配备了由PI开发的基于实时视频的瑞利光学系统，该系统被大量用于沉降速度和沉降平衡分析。快速采集瑞利系统允许高效应用高精度时间导数方法，也是由斯塔福德博士开发的，用于沉降速度分析，实现超离心光学灵敏度提高100-1000倍。e型已变得越来越难以维护，它是不太可能的，贝克曼仪器将继续支持他们后，他们在1995年底推出瑞利光学系统。多年来，bbi的许多研究人员以及许多外部合作者都从超离心设备中受益。AURL的持续可行性需要采购配备瑞利光学系统的新型XL-A。该仪器将由bri的七个主要研究小组共享，并将用于几个正在进行或计划进行的外部合作。其中一些研究人员过去曾受益于该设施。其他人要么目前或将来将使用超离心设备来研究各种蛋白质-蛋白质和蛋白质-核酸的相互作用。这些包括但不限于：P. and A. Wang， Caldesmon的结构、分子量以及磷酸化对其与肌动蛋白相互作用的影响；T. Tao，钙钙蛋白的结构和分子量；未来的研究将涉及钙钙蛋白与肌动蛋白相互作用的研究；Tao，肌钙蛋白I和肌钙蛋白C相互作用对钙的依赖性；陆瑞，肌凝蛋白调控和必需轻链与肌凝蛋白Sl头克隆调控域的相互作用；S.S. Lehrer， calponin和caldesmon与原肌凝蛋白的相互作用，以及各种原肌凝蛋白克隆片段端到端聚合特性的研究；H. Wohlrab，线粒体磷酸转运蛋白的性质及其亚基之间相互作用的研究；N. Sarkar，重组聚(A)聚合酶I和聚(A)聚合酶I的特性及其与引物RNA的相互作用；Z. Grabarek，钙调蛋白和肌钙蛋白c的钙依赖性构象变化及其与靶蛋白相互作用的钙依赖性研究；利用scFv免疫毒素和酸释放性抗白喉毒素（DT）单克隆抗体靶向c-erbB-2表位

This is a proposal to acquire a Beckman Instruments Optima Series XL-A Analytical Ultracentrifuge equipped with UV scanner and Rayleigh optical systems. This instrument will replace 2 heavily used Beckman Model-E's in the Analytical Ultracentrifuge Research Laboratory (AURL) at Boston Biomedical Research Institute (BBRI). One of the Model-E's is currently equipped with a real-time video-based Rayleigh optical system, developed by the PI, that is being used heavily in sedimentation velocity and sedimentation equilibrium analyses. The rapid acquisition Rayleigh system has allowed the efficient application of the high precision time derivative method, also developed by Dr. Stafford, for sedimentation velocity analysis achieving a 100-1000 fold increase in the sensitivity of the ultracentrifuge optics. The Model-E's have become increasingly difficult to maintain and it is unlikely that Beckman Instruments will continue to support them after they introduce the Rayleigh optical system in late 1995. Many researchers at BBRI as well as many outside collaborators have benefited from the ultracentrifuge facility over the years. The continued viability of the AURL requires the acquisition of a new XL-A equipped with Rayleigh optics. The instrument will be shared by seven major research groups at BBRI and will be used in several on-going or planned outside collaborations. Some of these researchers have benefited from the facility in the past. Others are either currently or will in the future use the ultracentrifuge facility to investigate various protein-protein and protein-nucleic acid interactions. These include but are not limited to: P. and A. Wang, structure, molecular weight of Caldesmon and the affect of phosphorylation on its interactions with actin; T. Tao, structure and molecular weight of calponin; future studies will involve studies of the interaction of calponin with actin; T Tao, calcium dependence of interactions between troponin I and troponin C; R Lu, interactions of myosin regulatory and essential light chains with the cloned regulatory domain of the myosin Sl head; S.S. Lehrer, interactions of calponin and caldesmon with tropomyosin, and studies of the end-to-end polymerization properties of various cloned fragments of tropomyosin; H. Wohlrab, investigation of properties of the mitochondrial phosphate transport protein and interactions between it subunits; N. Sarkar, characterization of recombinant poly (A) polymerase I and poly(A) polymerase I and their interactions with primer RNA; Z. Grabarek, investigation of calcium dependent conformational changes in calmodulin and troponin-C and the calcium dependence of their interactions with target proteins; V. Raso, Targeting c-erbB-2 Epitopes with scFv Immunotoxins and Acid-Releasable Anti-Diphtheria Toxin (DT) Monoclonal Antibodies.