Control of Executioner Caspases with an Allosteric Switch

用变构开关控制刽子手半胱天冬酶

• 批准号: 8255545
• 负责人: Jeanne Ann Hardy
• 金额: $ 24.96万
• 依托单位: UNIVERSITY OF MASSACHUSETTS AMHERST
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-07-01 至 2014-08-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8255545
• 关键词: Ablation; Active Sites; Adrenoleukodystrophy; Allosteric Site; Alzheimer' s Disease; Antineoplastic Agents; Apoptosis; Apoptotic; Benchmarking; Binding; Biochemical; Biological; Biological Process; Caspase; Caspase Inhibitor; Cell Death; Cells; Chemicals; Cleaved cell; Complex; Couples; Crystallography; Cysteine Protease; Data; Development; Development Plans; Diabetes Mellitus; Disease; Dissection; Family; Family member; Future; Genes; Genetic; Human; Individual; Intervention; Knock-out; Knockout Mice; Lead; Lesch-Nyhan Syndrome; Lysosomal Storage Diseases; Methods; Modeling; Myocardial Infarction; Names; Oculocerebrorenal Syndrome; Paper; Pathway interactions; Peroxisome Proliferator-Activated Receptors; Pharmaceutical Preparations; Phenotype; Play; Process; Property; Protein Family; Proteins; RNA Interference; Relative (related person); Research; Role; Specificity; Stroke; Structure; System; Systems Development; Technology; Time; biological systems; cancer cell; cancer therapy; caspase-3; caspase-6; caspase-7; clinical phenotype; design; directed evolution; human disease; improved; inhibitor/antagonist; interest; member; prevent; protein function; research study; small molecule; tool

DESCRIPTION (provided by applicant): Orthogonal control of protein function allows one protein from a family of related proteins to be inactivated so that its unique biological function can be assessed. We propose development of a new kind of orthogonal control, called allosteric switches, in the caspase family. Caspases are cysteine proteases that execute cell death by cleaving a discrete selection of target proteins, which ultimately leads to apoptosis. Caspases have a large central cavity that can be allosterically inhibited by small molecules. Caspases are believed to be good targets for treatment of cancer (caspase activators) or stroke, heart attack and Alzheimer's Disease (caspase inhibitors). Unfortunately the similarities in currently available small inhibitors that bind at the caspase active sites have not allowed unambiguous identification of the precise roles of the twelve different caspases. We will use a combination of directed evolution and computational protein design to introduce an allosteric switch in caspases-3, -6, and -7. When the small molecule effector we have selected or designed against binds to the evolved allosteric site, caspase activity will be inhibited. This switch will allow us to turn off one type of caspase at a time to determine what proteins are specifically cleaved by caspase-3 or -6 or -7. The cleaved proteins themselves are inherently interesting, because their cleavage is proapoptotic, meaning they lead to cell death. The ability to induce apoptosis is the hallmark of a useful cancer drug. Our studies may suggest which caspase should be targeted for treating which disease. Successful implementation of allosteric switches in these caspases will pave the way for allosteric switches in the other nine caspases and ultimately into other families of proteins where biological information is lacking. NON-TECHNICAL SUMMARY: The central problem in disease treatment is knowing which protein should be targeted to treat which disease. The allosteric switch technology we will develop in caspases allows just one type of protein to be turned off by a drug, to determine which disease could be effectively treated by targeting that particular protein. We focus on caspase proteins, which can cause cancer cells to die.

描述（由申请人提供）：蛋白质功能的正交控制允许相关蛋白质家族中的一种蛋白质失活，从而可以评估其独特的生物学功能。我们建议在半胱天冬酶家族中开发一种新型正交控制，称为变构开关。半胱天冬酶是半胱氨酸蛋白酶，通过切割离散选择的靶蛋白来执行细胞死亡，最终导致细胞凋亡。半胱天冬酶有一个大的中央空腔，可以被小分子变构抑制。 Caspase 被认为是治疗癌症（Caspase 激活剂）或中风、心脏病和阿尔茨海默病（Caspase 抑制剂）的良好靶标。不幸的是，目前可用的与 caspase 活性位点结合的小型抑制剂的相似性不允许明确识别 12 种不同 caspase 的精确作用。我们将结合定向进化和计算蛋白质设计在 caspase-3、-6 和 -7 中引入变构开关。当我们选择或设计的小分子效应物与进化的变构位点结合时，半胱天冬酶活性将受到抑制。这个开关将允许我们一次关闭一种类型的 caspase，以确定哪些蛋白质被 caspase-3 或 -6 或 -7 特异性切割。切割的蛋白质本身就很有趣，因为它们的切割是促凋亡的，这意味着它们会导致细胞死亡。诱导细胞凋亡的能力是有用的抗癌药物的标志。我们的研究可能表明应该针对哪种 caspase 来治疗哪种疾病。在这些半胱天冬酶中成功实施变构开关将为其他九种半胱天冬酶中的变构开关并最终进入缺乏生物信息的其他蛋白质家族铺平道路。 非技术摘要：疾病治疗的核心问题是知道应该针对哪种蛋白质来治疗哪种疾病。我们将在半胱天冬酶中开发的变构开关技术允许药物仅关闭一种类型的蛋白质，从而确定通过针对该特定蛋白质可以有效治疗哪种疾病。我们专注于半胱天冬酶蛋白，它可以导致癌细胞死亡。