Regulation of GRP94: Targeting Novel Cancer Therapeutics

GRP94 的调控：针对新型癌症疗法

• 批准号: 6550233
• 负责人: JEFFREY P BAKER
• 金额: $ 4.42万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-07-01 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6550233
• 关键词: adenosine diphosphate; adenosine triphosphate; affinity chromatography; antineoplastics; binding proteins; binding sites; calorimetry; cell line; chimeric proteins; dimer; drug discovery /isolation; endoplasmic reticulum; epidermal growth factor; flow cytometry; gel electrophoresis; growth factor receptors; membrane proteins; molecular chaperones; pharmacokinetics; point mutation; postdoctoral investigator; protein protein interaction; protein structure function; protein transport; stoichiometry; stress proteins

DESCRIPTION (provided by applicant): The mechanism of GRP94 regulation remains an open topic. GRP94 is a homodimer containing two identical ligand-binding sites. These ligand-binding sites are targets for the natural anti-tumor compounds geldanamycin (GA) and radicicol (RD) and also function in ATP/ADP binding. Adenosine and its analogues are known to bind GRP94 at a stoichiometry of 1 mol ligand: GRP94 dimer and such binding is regulated through negative cooperativity. We hypothesize that the negative cooperativity of ATP/ADP binding functions to provide a highly sensitive response to cell stress conditions that elicit a decrease in cellular ATP/ADP levels. GA and RD compete nucleotide binding and are able to bind GRP94 at a stoichiometry of 2-mol ligand: GRP94 dimer. We hypothesize that inhibitor ligands that target the adenosine nucleotide binding pocket of GRP94 and override negative cooperativity will function as a novel class of cancer chemotherapeutics. To examine this hypothesis, a broad array of GRP94 chimera will be studied to identify the region(s) of the molecule responsible for the negative cooperativity. In addition to negative cooperativity, there are likely to be other regions of the molecule critical to GRP94 function. To define such additional roles, individual GRP94 domains will be over-expressed and screened for dominant negative inhibitor activity. Together, the proposed analysis will lead to a greater understanding of GRP94 as a novel anti-cancer target.

描述（由申请人提供）：GRP94法规的机制仍然是一个公开主题。 GRP94是一种均匀的二聚体，包含两个相同的配体结合位点。这些配体结合位点是天然抗肿瘤化合物Geldanamycin（GA）和Radicicol（RD）的靶标，并且在ATP/ADP结合中也起作用。已知腺苷及其类似物在1 mol配体的化学计量法上结合GRP94：GRP94二聚体，这种结合通过负合作性调节。我们假设ATP/ADP结合函数的负合作性对细胞应力条件的高度敏感反应提供了降低的细胞应力条件，从而降低了细胞ATP/ADP水平。 GA和RD竞争核苷酸的结合，并能够在2-Mol配体的化学计量法上结合GRP94：GRP94二聚体。我们假设针对GRP94的腺苷核苷酸结合口袋和负合作的抑制剂配体将充当一类新型的癌症化学治疗剂。为了审查这一假设，将研究一系列的GRP94嵌合体，以识别负责负合作性的分子的区域。除了负合作性外，该分子的其他区域可能对GRP94功能至关重要。为了定义此类额外的角色，单个GRP94域将被过表达并筛选出主要的负面抑制剂活性。总之，提出的分析将使人们对GRP94作为一种新的抗癌目标有更深入的了解。