PRIMARY STRUCTURE ANALYSIS OF REGULATORY PROTEINS

自上次报告以来，分析技术不断改进， 已经扩展了内部氨基酸序列的实际下限 分析通过凝胶电泳分离的蛋白质， 50 pmol。 这允许分析几种新的蛋白质， 促进了以前研究的蛋白质的分析。 获得了足够的环GMP序列信息， 抑制磷酸二酯酶在上一份报告中描述， 克隆和表达。 大约300个残基的独特序列 已经获得了一种新的神经元钙调素结合蛋白。 尽管已经获得了足够的序列来解释超过60%的 该蛋白一级结构与其它蛋白没有明显同源性 哺乳动物蛋白质已被检测到。 一种展示紫外线的蛋白质 辐射诱导的DNA结合活性已被测序，允许 全长cDNA的分离。 在与腺苷酸环化酶（Z 01-BB-1）的纯化相关的研究期间， 07004-03），几种与毛喉素亲和支持物结合的蛋白质， 观察 其中三个已被确定为细胞骨架 微管蛋白、肌动蛋白和网格蛋白。 几种蛋白结合假定的第二信使Ap 5A也被发现。 通过序列分析鉴定为各种糖酵解酶， 这表明了一种新的监管水平的可能性， 流程. 这些技术的一般效用也被用来支持 在CBER内努力确定特定组件， 导致其不良影响的过敏材料 最终的目标是使过敏原的标准化 产品. 苹果和小麦抗原的一级序列 与过敏患者的血清进行比较， 未描述的蛋白质

Since the last report, continued improvements in analytical technology have extended the lower practical limit for internal amino acid sequence analysis of proteins separated by gel electrophoresis to the vicinity of 50 pmol. This has allowed analysis of several novel proteins and has facilitated analysis of proteins previously under study. Sufficient sequence information was obtained for the cyclic GMP- inhibited phosphodiesterase described in the last report to allow cloning and expression. Approximately 300 residues of unique sequence have been obtained for a novel neuronal calmodulin binding protein. Though sufficient sequence has been obtained to account for over 60% of the protein's primary structure, no significant homology with other mammalian proteins has been detected. A protein demonstrating UV irradiation-induced DNA binding activity has been sequenced, allowing isolation of the full-length cDNA. During studies related to purification of adenylyl cyclase (Z01-BB- 07004-03), several proteins binding to forskolin affinity supports were observed. Three of these have been identified as the cytoskeletal proteins tubulin, actin, and clathrin. Several proteins binding the putative second messenger Ap5A were also identified by sequence analysis as various glycolytic enzymes, indicating the possibility of a novel level of regulation of these processes. The general utility of these techniques has also been used to support efforts within CBER to identify the particular components within allergenic materials that are responsible for their untoward effects with the eventual goal of allowing better standardization of allergenic products. Primary sequence from apple and wheat antigens that react with sera from allergic patients has identified them as previously undescribed proteins.