FIRST CRYSTAL STRUCTURE OF PROTEIN KINASE

蛋白激酶的第一个晶体结构

• 批准号: 2465607
• 负责人: JANUSZ M SOWADSKI
• 金额: $ 22.09万
• 依托单位: TUFTS UNIVERSITY BOSTON
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-04-01 至 1999-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2465607
• 关键词: X ray crystallography; active sites; adenosine diphosphate; adenosine triphosphate; conformation; crystallization; enzyme inhibitors; enzyme mechanism; enzyme structure; enzyme substrate; enzyme substrate analog; enzyme substrate complex; isozymes; magnesium; manganese; nucleotide analog; phosphorylation; protein kinase A; staurosporine; structural biology

DESCRIPTION: How external growth signals are transmitted from the cell surface to the nucleus is one of the fundamental issues of cell biology. Research in the field of signal transduction has been motivated by the assumption that understanding of the signal pathways responsible for cell growth will yield insight into the uncontrolled growth seen in cancer. Recently, this assumption has been extensively confirmed by the discovery of the detailed molecular mechanism of cell cycle in which a key role is assigned to the protein kinase and its activators and inhibitors. The long-term goal is the rational design of inhibitors for protein kinases. This will incorporate co-crystallization of cAPK with metavanadate and nucleotide "caged" and peptide "caged" complexes, and co-crystallization of a staurosporin class of inhibitors, a derivative of which became a potent specific inhibitor of protein kinase C and EGFR kinase. Co-crystallization with the caged analogs, carried out in collaboration with Professor Roger Goody, is aimed at dynamic analysis of conformational changes of the catalytic core, namely the glycine-rich loop. Co-crystallization of staurosporin inhibitors, carried out in collaboration with Ciba-Geigy A.G., Basil, aims at enhancing the specificity of those inhibitors using crystallography and modeling. The current studies aim at the structural basis of phosphotransfer and substrate recognition of cAPK, the first protein kinase whose structure became known. The three specific aims are: (1) The analysis of conformational changes which the conserved catalytic core undergoes. (2) The structure of the transition state analog. (3) The mechanism of inhibition of catalysis through binding of a potent inhibitor - staurosporin. These aims will be accomplished using eight crystals of the recombinant unmyristylated catalytic subunit of protein kinase A and using the refined set of phases and methods of molecular replacement.

描述：外部生长信号如何从细胞传输 表面到细胞核是细胞生物学的基本问题之一。 信号转导领域的研究受到以下因素的推动 假设了解负责细胞的信号通路 生长将有助于深入了解癌症中不受控制的生长。 最近，这个假设被广泛证实 细胞周期的详细分子机制，其中关键作用是 分配给蛋白激酶及其激活剂和抑制剂。 这 长期目标是合理设计蛋白激酶抑制剂。 这将结合 cAPK 与偏钒酸盐的共结晶， 核苷酸“笼状”和肽“笼状”复合物，以及共结晶 星形孢菌素类抑制剂，其衍生物成为有效的 蛋白激酶C和EGFR激酶的特异性抑制剂。 共结晶 与笼中的类似物，与罗杰教授合作进行 Goody，旨在动态分析构象变化 催化核心，即富含甘氨酸的环。 共结晶 星形孢菌素抑制剂，与 Ciba-Geigy A.G. 合作进行， Basil，旨在增强这些抑制剂的特异性 晶体学和建模。 目前的研究主要针对结构 cAPK 磷酸转移和底物识别的基础，第一个 其结构已为人所知的蛋白激酶。 这三个具体目标是： (1) 保守催化的构象变化分析 核心经历。 (2)过渡态模拟的结构。 (3) 的 通过与有效抑制剂结合抑制催化的机制 - 星形孢菌素。 这些目标将使用八个晶体来实现 重组蛋白激酶A的未肉豆蔻酰化催化亚基并使用 分子置换的精确阶段和方法集。