CALCIUM ION TRANSPORT ATPASE AND ALTERED CALCIUM ION HOMEOSTASIS

细胞内Ca ~（2+）在许多细胞过程中起着重要作用 包括增殖、分化和肌肉收缩/关系。 他 肌质/内质网Ca 2+转运ATP酶（SERCA）是关键 参与调节细胞质Ca 2+和细胞内Ca 2+的酶 Ca2_ stores. 可能发生SERCA表达和/或功能的改变 在某些生理或病理生理条件下， 高血压和心力衰竭。 SERCA泵的特定抑制剂是 越来越多地用于研究：（1）细胞内Ca 2+稳态，（2） SERCA基因表达的调控，以及（3）结构-功能 与SERCA的关系。 我们开发了一个新的模型系统， 这些SERCA泵抑制剂，这构成了目前的基础， 应用程序. 特别是，我们在各种选择下开发了 策略，对抑制剂具有高度抗性的哺乳动物细胞系 特异性SERCA泵阻断剂毒胡萝卜素（TG）和2，5- 二（叔丁基）-1，4-氢醌（DBHQ）。 这导致某些 生物化学和功能的适应，相对于细胞的Ca 2 + 耐药细胞系中的稳态和SERCA泵。 在这 应用，我们提出了一系列详细的分子生物学研究， 存在于耐药细胞系中的SERCA泵，以便解剖 潜在的适应范围，可以发生在他们方面， 在这种特定的选择压力下表达和功能。 这些 研究将产生新的信息机制控制 改变（和潜在的病理性）Ca 2+稳态，这与 对于理解与异常相关的疾病状态也很重要 在Ca 2+稳态中。

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.