CIRCULAR DICHROISM SPECTROPOLARIMETER

这份共享仪器赠款申请旨在为以下项目提供资金： 购买圆二色光谱旋光仪， 处理和计算机控制能力。 使用本 该工具将由九个NIH受资助者共享。 它会让 现有的能力，检查二级结构的一个 多种蛋白质，并用于估计蛋白质的变化 在结合或取代金属离子时或在 通过体外诱变改变氨基酸序列。 的 离子结合蛋白包括蛇毒蛋白酶（Fox）， 钙调素（Kretsinger）和几种与 胞吐和囊泡融合（Creutz）。 待检蛋白质 突变后改变包括葡萄球菌核酸酶 （Benjamin），E.大肠杆菌维生素B12受体（Kadner）， 组织相容性抗原（Engelhard），src癌基因产物 （帕森斯），酵母组蛋白（史密斯），和水泡性口炎病毒 整合和外周膜蛋白（Rheuman和瓦格纳）。 这些研究的信息将是非常宝贵的， 单个氨基酸取代是否影响 蛋白质通过直接作用于改变的位点， 蛋白质结构的破坏或不稳定，或 子域。 因此，所请求的仪器将播放 在大量与主题无关的研究中发挥重要作用 项目，其中没有一个将以其他方式证明购买这种 设备. 我们希望能提供证据证明这一点的价值 基因工程表征中的仪器 proteins.

This request for a Shared Instrumentation Grant seeks funds for purchase of a circular dichroism spectropoloarimeter with data processing and computer control capabilities. Use of this instrument will be shared by nine NIH grantees. It will make available the capacity for examination of secondary structure of a variety of proteins and for estimation of the changes in protein conformation upon binding or substitution of metal ions or upon alteration of amino acid sequence by in vitro mutagenesis. The ion-binding proteins include snake venom proteases (Fox), calmodulin (Kretsinger), and several proteins implicated in exocytosis and vesicle fusion (Creutz). Proteins to be examined after mutational alteration include staphylococcal nuclease (Benjamin), the E. coli vitamin B12 receptor (Kadner), histocompatibility antigens (Engelhard), the src oncogene product (Parsons), yeast histones (Smith), and the vesicular stomatits virus integral and peripheral membrane proteins (Pal and Wagner). Information from these studies will be invaluable in showing whether single amino acid substitutions influence the activity of that protein through direct action at the altered site or by disruption or labilization of the structure of the protein or a subdomain. Thus, the requested instrumentation will play an important role in a large number of topically unrelated research projects, none of which would otherwise justify purchase of such equipment. We hope to provide evidence showing the value of this instrumentation in the characterization of genetically engineered proteins.