ACTIVE SITES OF GLYCYL-TRNA SYNTHETASE--EXTRACELLULAR SEA URCHIN EMBRYO PROTEINS

甘氨酰-TRNA合成酶活性位点--海胆胚胎胞外蛋白

• 批准号: 6240369
• 负责人: GLENN M NAGEL
• 金额: $ 4.39万
• 依托单位: CALIFORNIA STATE UNIVERSITY FULLERTON
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-02-01 至 1998-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6240369
• 关键词: active sites; alternatives to animals in research; aminoacid tRNA ligase; chemical binding; embryo /fetus protein; enzyme activity; enzyme structure; extracellular matrix; fertilization; hyaline substance; protein biosynthesis; protein sequence; radionuclides; sea urchins

Students will work in two broad research areas. In the first project, structure and functions of the enzyme glycyl-tRNA synthetase will be investigated in order to provide information concerning basic molecular mechanisms of protein synthesis. This work will concentrate particularly on the role placed by dissimilar subunits in the recognition and activation of tRNA and ATP. Affinity labeling reagents will be employed to probe the enzyme's recognition sites and modified peptides isolated to locate essential amino acid residues in the active site of the enzyme. Subsequent studies will be directed at a closely related enzyme, phenylalanyl-tRNA synthetase. Computer analysis will compare amino acid sequences so identified with those of other aminoacyl -tRNA synthetases to better understand how these enzymes as a group accomplish their essential role in vivo. In the second project, the proteins forming the extracellular matrix of sea urchin embryos following fertilization will be investigated. By purifying the protein components of the hyaline layer and the fertilization envelope, we hope to provide an understanding of the molecular mechanisms of cellular organization in the embryo and the manner in which the block to polyspermic fertilization is accomplished. A major goal of this work is to provide information regarding the amino acid sequence of three proteins already isolated by students in the laboratory. This work aims to correlate amino acid sequences of the proteins with DNA sequences of hyalin-associated proteins presently being completed in other laboratories. Together, these data can serve to understand how the three proteins are related to one another and to other calcium-binding and extracellular matrix protein as well as how these proteins are regulated developmentally and how they perform their essential biological role. The primary goal of this project is to provide a scientific environment which gives these students the opportunity to succeed as young scientists and an atmosphere which supports and encourages academic involvement and achievement. Students form our laboratory have established a strong record of accomplishment. Of the fifty-nine students who have completed projects since 1980, 44% have entered graduate or professional programs.

学生将在两个广泛的研究领域工作。 在第一个项目中， 甘氨酰-tRNA合成酶的结构和功能将被 为了提供有关基本分子的信息， 蛋白质合成的机制。 这项工作将特别集中 不同亚单位在识别中的作用， 激活tRNA和ATP。 将使用亲和标记试剂 以探测酶的识别位点和分离的修饰肽， 将必需氨基酸残基定位在酶的活性部位。 随后的研究将针对一种密切相关的酶， 苯丙氨酰-tRNA合成酶。计算机分析将比较氨基酸 与其他氨酰-tRNA合成酶的序列相同， 更好地了解这些酶作为一个群体是如何完成其基本功能的。 在体内的作用。 在第二个项目中，形成细胞外基质的蛋白质 将研究受精后的海胆胚胎。 通过 纯化透明层的蛋白质组分， 受精信封，我们希望提供一个理解的 胚胎中细胞组织的分子机制以及 在这个过程中完成了对多精受精的阻断 一个主要 这项工作的目的是提供有关氨基酸的信息， 学生们已经在实验室里分离出了三种蛋白质的序列。 这项工作的目的是将蛋白质的氨基酸序列与DNA 透明蛋白相关蛋白的序列目前正在其他领域完成， laboratories. 总之，这些数据可以用来了解这三个 蛋白质彼此相关，并与其他钙结合， 细胞外基质蛋白以及这些蛋白质是如何调节的 以及它们如何发挥其重要的生物学作用。 的 该项目的主要目标是提供一个科学的环境， 使这些学生有机会成为成功的年轻科学家， 支持和鼓励学术参与的氛围， 成就 我们实验室的学生已经建立了一个强大的 成就的记录。 在59名已经完成 自1980年以来，44%的人进入了研究生或专业课程。