CALCIUM ION TRANSPORT ATPASE AND ALTERED CALCIUM ION HOMEOSTASIS

钙离子转运ATP酶和改变的钙离子稳态

• 批准号: 6241717
• 负责人: Arif Hussain
• 金额: $ 22.69万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 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 1998-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6241717
• 关键词: calcium flux; calcium metabolism; calcium transporting ATPase; cell line; enzyme activity; enzyme inhibitors; enzyme mechanism; enzyme structure; gene expression; homeostasis; hydroquinones; protein structure function; sarcoplasmic reticulum; thapsigargin

Intracellular Ca2+ plays a fundamental role in many cellular processes including proliferation, differentiation, and muscle contraction/relation. he sarcoplasmic/endoplasmic reticulum Ca2+ transport ATPase (SERCAs) are key enzymes involved in the regulation of cytoplasmic Ca2+ and intracellular Ca2_ stores. Alterations in SERCA expression and/or function can occur under certain physiological or pathophysiological conditions including hypertension and heart failure. Specific inhibitors of the SERCA pump are increasingly being used to study: (1) intracellular Ca2+ homeostasis, (2) regulation of SERCA gene expression, and (3) structure-function relationships within SERCA. We have developed a novel model system based on these SERCA pump inhibitors, which forms the basis for the current application. In particular, we have developed, under various selection strategies, mammalian cell lines that are highly resistant to the inhibitor effects of the specific SERCA pump blockers thapsigargin (TG) and 2,5- di(tert-butyl)-1, 4 hydroquinone (DBHQ). This results in certain biochemical and functional adaptations with respect to cellular Ca2+ homeostasis and SERCA pumps in the resistant cell lines. In this application, we propose a series of detailed molecular biological studies o the SERCA pumps present in the resistant cell lines in order to dissect the potential range of adaptations that can occur within them with respect to expression and function under such specific selective pressure. These studies will yield novel information regarding mechanisms controlling altered (and potentially pathologic) Ca2+ homeostasis, which are relevant and important to understanding disease states associated with abnormalities in Ca2+ homeostasis.

细胞内 Ca2+ 在许多细胞过程中发挥着基础作用 包括增殖、分化和肌肉收缩/关系。 他 肌浆/内质网 Ca2+ 转运 ATP 酶 (SERCA) 是关键 参与细胞质 Ca2+ 和细胞内 Ca2+ 调节的酶 Ca2_ 储存。 SERCA 表达和/或功能可能发生改变 在某些生理或病理生理条件下，包括 高血压和心力衰竭。 SERCA 泵的具体抑制剂是 越来越多地用于研究：(1) 细胞内 Ca2+ 稳态，(2) SERCA 基因表达的调控，以及 (3) 结构-功能 SERCA 内部的关系。 我们开发了一种新颖的模型系统 这些 SERCA 泵抑制剂，构成了当前的基础 应用。 特别是，我们在各种选择下开发了 策略，对抑制剂高度耐药的哺乳动物细胞系 特定 SERCA 泵阻滞剂毒胡萝卜素 (TG) 和 2,5- 的作用 二(叔丁基)-1, 4 对苯二酚 (DBHQ)。 这会导致某些 细胞 Ca2+ 的生化和功能适应 耐药细胞系中的稳态和 SERCA 泵。 在这个 应用中，我们提出了一系列详细的分子生物学研究 SERCA 泵存在于耐药细胞系中，以便剖析 它们内部可能发生的潜在适应范围 在这种特定的选择压力下的表达和功能。 这些 研究将产生有关控制机制的新信息 改变（并且可能是病理性的）Ca2+稳态，这是相关的 对于了解与异常相关的疾病状态很重要 Ca2+ 稳态。