ANESTHETIC EFFECTS ON SARCOPLASMIC RETICULUM CA++ ATPASE

描述：（改编自申请人摘要）初步 根据申请人的实验，已经发现全身和局部麻醉剂 影响Ca-ATP酶的活性和寡聚状态， 骨骼肌和心肌的肌浆网（SR）。 这些 结果是一致的解释，四分体和较大的 低聚物的活性比Ca-ATP酶的单体和二聚体低得多。 的 长期目标是阐明这些的分子机制 麻醉剂对Ca-ATP酶的影响，并了解其相关性 全身麻醉药的心肌抑制作用， 局部麻醉药的抗惊厥作用。 将标记Ca-ATPase 用共价磷光探针，时间分辨磷光 将使用光谱法来估计Ca-ATP酶的寡聚状态 在心脏和骨骼肌的SR膜中。 将军的影响 和局麻药对Ca-ATP酶寡聚状态和活性的影响 将确定心脏和骨骼SR。 AT-1 SR的实验 细胞，一种缺乏受磷蛋白的心房肿瘤细胞系，将有助于 确定受磷蛋白是否是导致 心脏和骨骼肌SR Ca-ATP酶对全身麻醉药的反应。 的 申请人将确定麻醉剂对以下反应的影响： 心肌Ca-ATP酶的寡聚状态和活性与其两个主要 生理调节剂，钙和磷蛋白磷酸化，副 亦然 在骨骼和心脏SR中，他们将询问 局部麻醉剂促进大寡聚体形成的作用 Ca-ATP酶依赖于带正电荷的季铵部分， 大多数局部麻醉剂。 麻醉药对麻醉反应的影响 心脏和骨骼Ca-ATPase改变pH值，反之亦然，将是 研究了 这些文献记载了深刻而鲜为人知的 pH值对心肌肌浆网Ca-ATP酶对吸入氧自由基反应的影响 麻醉剂，拟议的实验旨在解释。 原子 力显微镜将提供一种独立的方法来证实或反驳 麻醉剂诱导Ca-ATP酶寡聚体状态的变化， 由先前的时间分辨磷光结果所示 谱

DESCRIPTION: (Adapted from the applicant's abstract) In preliminary experiments of the applicant, general and local anesthetics have been found to influence the activity and the oligomeric state of the Ca-ATPase of sarcoplasmic reticulum (SR) of both skeletal and cardiac muscle. These results are consistent with the interpretation that tetrameres and larger oligomers are much less active than monomers and dimers of Ca-ATPase. The long term objective is to elucidate the molecular mechanisms of these effects of anesthetics on the Ca-ATPase and to understand their relevance to the myocardial depressive effects of general anesthetics and the antiarrhythmic effects of local anesthetics. Ca-ATPase will be labeled with a covalent phosphorescent probe, and time resolved phosphorence spectroscopy will be used to estimate the oligomeric state of the Ca-ATPase in SR membranes of cardiac and skeletal muscle. The influences of general and local anesthetics on the oligomeric state and activity of Ca-ATPase of cardiac and skeletal SR will be determined. Experiments with SR from AT-1 cells, an atrial tumor cell line deficient in phospholamban, will help to determine whether phospholamban is responsible for the very different responses of cardiac and skeletal SR Ca-ATPase to general anesthetics. The applicants will determine the influence of anesthetics on the responses of the oligomeric state and activity of cardiac Ca-ATPase to its two major physiologic regulators, Ca and phosphorylation of phospholamban, and vice versa. In both skeletal and cardiac SR, they will inquire whether the effect of local anesthetics to promote the formation of large oligomers of Ca-ATPase depends upon the positively charged quaternary ammonium moiety of most local anesthetics. The influence of anesthetics on the response of cardiac and skeletal Ca-ATPase to altered pH, and vice versa, will be investigated. The literature documents profound and poorly understood influences of pH on the responses of cardiac SR Ca-ATPase to inhalational anesthetics that the proposed experiments are designed to explain. Atomic force microscopy will provide an independent method to confirm or refute the anesthetic induced changes in oligomeric state of Ca-ATPase that are indicated by the previous results with time resolved phosphorescence spectroscopy.