Structural Studies of the Tumor M2 Isoform of Pyruvate Kinase

丙酮酸激酶肿瘤 M2 亚型的结构研究

• 批准号: 8619289
• 负责人: GREGORY DEAN BOWMAN
• 金额: $ 17.62万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-01-01 至 2015-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8619289
• 关键词: Adult; Affect; Allosteric Regulation; Amino Acids; Anabolism; Binding; Blood; Brain; Breast; Cancer Detection; Cancer cell line; Cancerous; Cell Proliferation; Cell Survival; Cell division; Cells; Characteristics; Complex; Detection; Development; Diagnosis; Embryo; Environment; Enzymes; Fructose; Glucose; Goals; Growth; Histone H1; Histone H1(s); Histone H3; Ligands; Link; Liver; Lung; Malignant Neoplasms; Metabolic; Metabolism; Molecular Conformation; N-terminal; Nucleic Acids; Nutrient; Peptides; Phospholipids; Phosphorylation; Phosphotransferases; Phosphotyrosine; Physiology; Protein Isoforms; Protein Kinase; Proteins; Purines; Pyruvate Kinase; Regulation; Research; Role; STAT3 gene; Specificity; Structure; Substrate Specificity; Therapeutic; Threonine; Tissues; Tyrosine; Work; X-Ray Crystallography; Xenograft procedure; aerobic glycolysis; base; cancer cell; cancer diagnosis; cancer prevention; cancer therapy; cancer type; cdc Genes; cell growth; dimer; insight; knock-down; novel therapeutics; programs; public health relevance; purine; small molecule; tumor; uptake

Cancerous cells maintain an altered metabolic state that dramatically increases the flux of nutrients into synthesizing cellular building blocks, which is essential for rapid cell growth and division. A key enzyme required for this proliferative metabolism is the M2 isoform of pyruvate kinase, called PKM2. PKM2 is found predominantly as a dimer and highly expressed in a multitude of cancer types, with a clear correlation between expression levels and tumor aggressiveness. Knock-down of PKM2 or substitution with the adult PKM1 isoform has been shown to reduce cancer cell proliferation, making PKM2 an important target for better understanding the development, diagnosis, and treatment of cancer. Recent work has revealed new and unexpected activities and interactions of PKM2 that are important for promoting cell proliferation. PKM2 binds to and is stimulated by SAICAR, a purine biosynthesis intermediate, and this interaction is important for cancer cell survival in nutrient-poor environments. PKM2 has also been shown to bind to phosphotyrosine-containing proteins, which stabilize PKM2 in a dimeric state. Interestingly, this dimeric state of PKM2 has been shown to have protein kinase activity, inducing altered transcriptional programs by phosphorylating targets such as histone H1 and H3, prothymosin ¿, and STAT3. This proposal aims to uncover the structural basis of PKM2 regulation and substrate specificity. We will use X-ray crystallography to capture PKM2 in complex with SAICAR and phosphorylation targets to reveal how these ligands allosterically influence PKM2 structure and function. The results of this research will help provide a structural framework for understanding how PKM2 participates in stimulating cell proliferation in normal and diseased cells.

癌细胞维持改变的代谢状态，可显着增加 营养成合成细胞构建基块，这对于快速细胞生长和分裂至关重要。一个 这种增殖代谢所需的关键酶是丙酮酸激酶的M2同工型，称为PKM2。 PKM2主要是二聚体，并且在多种癌症类型中高度表达， 表达水平与肿瘤攻击性之间的明显相关性。击倒PKM2或 已证明用成年PKM1同工型取代可以减少癌细胞的增殖，从而使 PKM2是更好地了解癌症的发育，诊断和治疗的重要目标。 最近的工作揭示了PKM2的新的和意外的活动和互动 对于促进细胞增殖很重要。 PKM2与Saicar结合并刺激 生物合成中间体，这种相互作用对于营养贫困的癌细胞存活很重要 环境。 PKM2也已显示与含磷酸酪氨酸的蛋白结合，该蛋白 稳定PKM2处于二聚体状态。有趣的是，已证明这种二聚体状态具有 蛋白激酶活性，通过磷酸化靶标（例如 Hisstone H1和H3，螺杆菌素€和STAT3。 该建议旨在揭示PKM2调节和底物特异性的结构基础。 我们将使用X射线晶体学来捕获与SAICAR和磷酸化目标的复合物中的PKM2 揭示这些配体如何变构影响PKM2的结构和功能。结果的结果 研究将有助于提供一个结构框架，以了解PKM2如何参与刺激 正常细胞和患病细胞中的细胞增殖。