3D-STRUCTURE OF THE CAMP-DEPENDENT PROTEIN KINASE

CAMP 依赖性蛋白激酶的 3D 结构

• 批准号: 3304084
• 负责人: Jeff Kuret
• 金额: $ 19.34万
• 依托单位: COLD SPRING HARBOR LABORATORY
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-05-01 至 1994-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3304084
• 关键词: Saccharomyces cerevisiae; X ray crystallography; active sites; chimeric proteins; computer simulation; conformation; enzyme inhibitors; enzyme structure; enzyme substrate; enzyme substrate analog; enzyme substrate complex; fungal genetics; isomorphous substitution; isozymes; molecular dynamics; physical model; protein engineering; protein folding; protein kinase; protein kinase A

Protein kinases are key regulatory molecules that participate in many biochemical pathways. As catalysts, these enzymes can amplify intra- or extracellular signals. By virtue of their broad yet limited substrate specificity, they are able to regulate more than one substrate protein at a time, coordinating intracellular metabolism. Understanding how protein kinases achieve the specificity necessary to regulate cell processes is an important problem in biology, and is the long-term goal of this proposal. Specifically, a detailed structural study of TPK1, the cAMP-dependent protein kinase catalytic subunit of Saccharomyces cerevisiae, is proposed. Existing crystals of TPK1 that diffract to 2.2 A resolution will be derivatized with heavy atom compounds, solving the crystallographic phase problem, and yielding its three-dimensional structure. The structure will reveal the location of key residues previously demonstrated to be involved in substrate recognition and catalysis, and reveal the overall folding of a protein kinase domain. This work will be extended by predicting the structure of other protein kinases that differ from TPK1 in substrate specificity using comparative modeling methods. The resulting prediction of protein kinase substrate selectively will be tested by in vitro mutagenesis. Chimeric kinases will be synthesized and characterized to relate the structural features predicted by computer modeling to enzymatic activity. A complete understanding of protein kinase structure will speed the development of specific inhibitors potentially useful in the treatment of neoplastic disease.

蛋白激酶是参与许多参与的关键调节分子 生化途径。 作为催化剂，这些酶可以扩增或 细胞外信号。 凭借其广泛但有限的底物 特异性，他们能够在A处调节多个底物蛋白 时间，协调细胞内代谢。 了解蛋白质 激酶达到调节细胞过程所需的特异性是 生物学中的重要问题，是该提议的长期目标。 具体而言，TPK1的详细结构研究，依赖营地 提出了酿酒酵母的蛋白激酶催化亚基。 衍射到2.2的TPK1的现有晶体将是一个分辨率 用重原子化合物衍生化，求解晶体学阶段 问题，并产生其三维结构。 结构将 揭示先前证明涉及的关键残留物的位置 在底物识别和催化中，并揭示了A的整体折叠 蛋白激酶结构域。 通过预测 其他蛋白激酶与底物中TPK1不同的结构 使用比较建模方法的特异性。 由此产生的预测 蛋白激酶底物的选择性将通过体外测试 诱变。 嵌合激酶将合成并表征为 将计算机建模预测的结构特征与酶促 活动。 对蛋白激酶结构的完全了解将加快 特定抑制剂的发展可能在治疗中有用 肿瘤疾病。