MECHANISTIC STUDIES OF AICAR TRANSFORMYLASE

AICAR转化酶的机理研究

• 批准号: 2377286
• 负责人: CRAIG E. CAMERON
• 金额: $ 10.49万
• 依托单位: PENNSYLVANIA STATE UNIVERSITY, THE
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-09-01 至 2002-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2377286
• 关键词: DNA footprinting; Escherichia coli; Saccharomyces cerevisiae; catalyst; enzyme mechanism; enzyme structure; enzyme substrate; genetic manipulation; genetic strain; hematopoietic stem cells; hydroxymethyltransferases; methotrexate; neoplasm /cancer chemotherapy; pharmacokinetics; polymerase chain reaction; site directed mutagenesis; yeast two hybrid system

DESCRIPTION (Applicant's Description): The long-term objectives of this proposal are to develop further 5-aminoimidazole-4-carboxamide ribotide transformylase/inosine monophosphate cyclohydrolase (ATIC) as a c h e m otherapeutic target for the treatment of cancer and to create methotrexate-resistant variants of ATIC suitable for use in hematopoietic stem c e ll support thereby expanding the use of methotrexate in high-dose chemotherapy protocols. The specific aims of this proposal are as follows: (1) to use pre-steady-state kinetic methods to define the minimal kinetic schemes for the two reactions catalyzed by ATIC; (2) to identify the regions of ATIC responsible for dimerization and substrate binding using approaches such as site-directed mutagenesis and protein footprinting; and (3) to develop a genetic screen in yeast to identify methotrexate-resistant variants of ATIC.

描述（申请人的描述）：本申请的长期目标 建议进一步开发5-氨基咪唑-4-甲酰胺核苷酸 转化酶/肌苷酸环化水解酶（ATIC）作为一种酶， 治疗癌症的新靶点， 适用于造血干细胞移植的ATIC的甲氨蝶呤抗性变体 干细胞支持，从而扩大了甲氨蝶呤在高剂量 化疗方案。 这项建议的具体目标如下： (1)使用预稳态动力学方法来定义最小动力学 （2）确定ATIC催化的反应区域 负责二聚化和底物结合的ATIC 例如定点诱变和蛋白质足迹法;和（3） 在酵母中进行遗传筛选，以确定对甲氨蝶呤耐药的 ATIC的变体。