Regulation of H+/Ca2+ exchange by Vcx1p in budding yeast

芽殖酵母中 Vcx1p 对 H /Ca2 交换的调节

• 批准号: 6989496
• 负责人: Veronica A. Segarra
• 金额: $ 4.21万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-15 至 2007-09-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6989496
• 关键词: Saccharomyces cerevisiae; affinity chromatography; calcineurin; calcium metabolism; fungal genetics; fungal proteins; gene expression; genetic regulation; immunoprecipitation; mass spectrometry; membrane transport proteins; mutant; phenotype; phosphorylation; predoctoral investigator; protein protein interaction; protein structure function; radiotracer; site directed mutagenesis

DESCRIPTION (provided by applicant): Vcx1p is a vacuolar Ca2+/H+ exchanger that transports Ca2+ into the vacuole with the energy provided by the H+ gradient in Saccharomyces cerevisiae. In the absence of Vcx1p activity, elevated levels of cytosolic free Ca2+ are prolonged, suggesting that Vcx1p plays a major role in cytosolic Ca2+ regulation. Activation of calcineurin, a phosphatase, results in the inhibition of Vcx1p. However, the molecular events downstream of calcineurin activation that result in Vcx1p inhibition are not known. This proposal presents the experiments by which I will: identify the residues of Vcx1 required for its regulation, determine whether Vcx1 activity is directly modulated by phosphorylation, identify the molecule(s) that interacts with Vcx1 and regulate its activity, and in-vitro reconstitute Vcx1 activity in order to kinetically characterize its regulation.

描述（由申请人提供）：Vcx1p是一种液泡Ca2+/H+交换剂，利用酿酒酵母中H+梯度提供的能量将Ca2+输送到液泡中。在缺乏Vcx1p活性的情况下，细胞质游离Ca2+水平升高的时间延长，表明Vcx1p在细胞质Ca2+调节中起主要作用。磷酸酶钙调磷酸酶的激活导致Vcx1p的抑制。然而，钙调磷酸酶激活下游导致Vcx1p抑制的分子事件尚不清楚。本提案提出了我将通过以下实验：鉴定其调节所需的Vcx1残基，确定Vcx1活性是否由磷酸化直接调节，鉴定与Vcx1相互作用并调节其活性的分子，并在体外重建Vcx1活性，以便动力学表征其调节。