Roles and mechanisms of Slo and Slack Channels in Brain

Slo 和 Slack 通道在大脑中的作用和机制

• 批准号: 6369473
• 负责人: LEONARD K KACZMAREK
• 金额: $ 147.41万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-04 至 2006-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6369473
• 关键词: neurobiology; potassium channel; protein structure function

DESCRIPTION (provided by applicant): A class of calcium-activated potassium channels termed BK(Ca) channels are enriched at presynaptic neurotransmitter release sites within the brain, and are believed to control both the amount and timing of neurotransmitter release. BK(Ca) channels are encoded by the Slo gene. Recent evidence suggests that a related gene, Slack, also contributes to BK(Ca)-like channels in neurons, and that some BK(Ca) channels may be comprised of heteromultimers of Slo and Slack channels subunits. The experiments in this proposal will characterize the molecular, biophysical and structural properties of BK(Ca) channels in both the somata and presynaptic terminals of native neurons, and in transfected cells. The role of the Slo and Slack channel subunits in normal synaptic transmission, and the mechanisms of their response to hypoxia, will be determined by patch clamp measurements coupled with genetic knockout approaches. The structure and function of the large carboxy-terminal regulatory region of the Slo channel subunit with its calcium-binding sites, and its interaction with other proteins, including the Slack subunit, will be determined by biophysical and biochemical measurements and by X-ray crystallographic methods. Finally the structure of intact BK(Ca) channels will be determined by cryo-electronmicroscopy. Knowledge of the molecular properties and regulation of BK(Ca) channels in normal and hypoxic neurons, together with structural determinations of their interactions with both drugs and natural ligands, will be key in the development of more effective treatments for disorders such as epilepsy and stroke.

描述（由申请人提供）：一类钙激活剂 称为 BK(Ca) 通道的钾通道在突触前富集 大脑内的神经递质释放位点，被认为可以控制 神经递质释放的量和时间。 BK(Ca) 通道是 由 Slo 基因编码。最近的证据表明，一个相关的基因 Slack 还有助于神经元中的 BK(Ca) 样通道，并且某些 BK(Ca) 通道可以由 Slo 和 Slack 通道的异多聚体组成 亚单位。该提案中的实验将表征分子， 体细胞和体细胞 BK(Ca) 通道的生物物理和结构特性 天然神经元和转染细胞的突触前末端。的作用 正常突触传递中的 Slo 和 Slack 通道亚基，以及 它们对缺氧的反应机制将通过膜片钳来确定 测量与基因敲除方法相结合。结构和 Slo 通道大羧基末端调节区的功能 亚基及其钙结合位点及其与其他亚基的相互作用 蛋白质，包括 Slack 亚基，将通过生物物理和 生化测量和 X 射线晶体学方法。最后 完整 BK(Ca) 通道的结构将由下式确定 冷冻电子显微镜。了解分子特性和调控 正常和缺氧神经元中 BK(Ca) 通道的变化，以及结构 确定它们与药物和天然配体的相互作用，将 是开发更有效的疾病治疗方法的关键，例如 癫痫和中风。