BIOCHEMICAL BASIS FOR THE CONTROL OF THE ALKALI METAL/H

控制碱金属/H 的生化基础

• 批准号: 2749765
• 负责人: ALEJANDRO ORTIZ
• 金额: $ 1.72万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-08-01 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/2749765
• 关键词: SDS polyacrylamide gel electrophoresis; Urodela; acid base balance; alkali metals; aminoacid; antiport; cell morphology; coulometry; erythrocytes; flame photometry; hydrogen potassium exchanging ATPase; immunoprecipitation; ion transport; membrane transport proteins; osmosis; phosphatase inhibitor; phosphorylation; sodium potassium exchanging ATPase

In Amphiuma red blood cells, the Na/H and K/H antiports mediate volume regulation after shrinkage and swelling, respectively. These ionic flux routes play a crucial role in viability of certain tumor cells and are directly responsible for cell injury in cardiomyocites during hypoxic episodes. However, the basis for their activation and inactivation remains obscure. We hypothesize that antiport protein phosphorylation status determines activity, while phosphorylation pattern dictates ion selectivity. To test this hypothesis we will monitor transporter activity in conjunction with biochemical studies designed to evaluate transporter phosphorylation state in order to address the following specific aims: 1) Determine the effect of the phosphatase inhibitor Calyculin A in the ionic fluxes through the Na/H and K/H exchange. 2) Perform experiments to determine the time course for activation and inactivation of the Alkali Metal/H exchanger during hyperosmotic and hyposmotic perturbations and fit the data to a minimal two state model (active and inactive) proposed by Jennings, M.L. and Al-Rohil, N. (1990). 3) Determine which antiporter amino acid residues are phosphorylated or dephosphorylated in response to shrinkage or swelling.

在Amphiuma红细胞中，Na/H和K/H的抗原介导 收缩和肿胀后的体积调节。 这些 离子通量路线在某些肿瘤细胞的生存力中起着至关重要的作用 并直接负责心肌损伤 低氧发作。 但是，它们激活的基础 失活仍然晦涩。 我们假设该抗蛋白质 磷酸化状态决定活性，而磷酸化状态 模式决定了离子的选择性。 为了检验该假设，我们将监视 转运蛋白活性与旨在 评估转运蛋白磷酸化状态，以解决 以下特定目的：1）确定磷酸酶的作用 通过Na/H和K/H的离子通量中的抑制剂钙符蛋白A 交换。 2）执行实验以确定时间课程 碱金属/H交换器的激活和灭活 过度渗透和低渗性扰动，并将数据拟合到A 詹宁斯提出的最小两个状态模型（主动和不活动）， M.L.和Al-Rohil，N。（1990）。 3）确定哪个抗腹剂 氨基酸残基被磷酸化或去磷酸化 对收缩或肿胀的反应。

项目成果

Activation of Na+/H+ and K+/H+ exchange by calyculin A in Amphiuma tridactylum red blood cells: implications for the control of volume-induced ion flux activity.

Amphiuma tridactylum 红细胞中花萼蛋白 A 激活 Na /H 和 K /H 交换：对控制体积诱导离子通量活性的影响。

• DOI: 10.1152/ajpcell.00160.2008
• 发表时间: 2008
• 期刊: American journal of physiology. Cell physiology
• 作者: Ortiz-Acevedo,Alejandro;Rigor,RobertR;Maldonado,HectorM;Cala,PeterM
• 通讯作者: Cala,PeterM