PRIMARY STRUCTURE ANALYSIS OF REGULATORY PROTEINS

调节蛋白的一级结构分析

• 批准号: 3792436
• 负责人: M MOOS
• 金额: --
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3792436
• 关键词: DNA binding protein; binding proteins; calmodulin; cyclic GMP; cytoskeletal proteins; forskolin; gel electrophoresis; glycoproteins; phosphoproteins; protein sequence; protein structure; radiation genetics; transcription factor; ultraviolet radiation

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.

自上次报告以来，分析技术的持续改进 延长了体内氨基酸序列的实用下限 凝胶电泳法分离的蛋白质及其附近的分析 50pmol.这使得分析几种新的蛋白质成为可能，并已经 促进了对先前正在研究的蛋白质的分析。 获得了足够的序列信息，用于环GMP- 抑制上次报告中描述的磷酸二酯酶，以允许 克隆和表达。大约300个唯一序列的残基 已经获得了一种新的神经元钙调蛋白结合蛋白。 虽然已经获得了足够的序列来解释超过60%的 蛋白质的一级结构，与其他蛋白质没有明显的同源性 哺乳动物的蛋白质已被检测到。一种显示紫外线的蛋白质 已经对辐射诱导的DNA结合活性进行了测序，从而 全长c DNA的分离。 腺酰环化酶(Z01-BB-)的纯化研究 07004-03)，与Forsklin亲和载体结合的几种蛋白质 观察到的。其中三种已被确认为细胞骨架 微管蛋白、肌动蛋白和网状蛋白。 与推测的第二信使Ap5A结合的几种蛋白质也被 经序列分析鉴定为多种糖酵解酶， 表明有可能对这些产品进行新的监管 流程。 这些技术的一般用途也被用来支持 在CBER内努力确定其中的特定组件 导致其不良反应的过敏性物质 最终目标是允许更好地标准化过敏原 产品。苹果和小麦抗原反应的一级序列 从过敏症患者的血清中鉴定出他们之前 未知蛋白质。