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

基本信息

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
项目类别：
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

项目摘要

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.

癌细胞维持一种代谢状态，显着增加癌细胞的通量营养物质合成细胞构件，这对于细胞快速生长和分裂至关重要。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 主要以二聚体形式存在，在多种癌症类型中高度表达， PKM2 或 PKM2 的表达水平与肿瘤侵袭性的敲低之间存在明确的相关性。已证明用成人 PKM1 亚型替代可以减少癌细胞增殖，使得 PKM2 是更好地了解癌症发展、诊断和治疗的重要靶点。最近的工作揭示了 PKM2 新的、意想不到的活动和相互作用， PKM2 与 SAICAR（一种嘌呤）结合并受其刺激。生物合成中间体，这种相互作用对于营养贫乏的癌细胞生存很重要 PKM2 还被证明可以与含有磷酸酪氨酸的蛋白质结合。 PKM2 处于二聚体状态，PKM2 的这种二聚体状态已被证明具有支持性。蛋白激酶活性，通过磷酸化靶标诱导转录程序，例如组蛋白 H1 和 H3、胸腺肽原 ¿和 STAT3。该提案旨在揭示 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 和磷酸化靶标复合的 PKM2 揭示这些配体如何变构影响 PKM2 的结构和功能。研究将有助于提供一个结构框架来理解 PKM2 如何参与刺激正常细胞和患病细胞的细胞增殖。