X-RAY STRUCTURES OF PENTAMERIC ION CHANNELS IN THE ABSENCE AND PRESENCE OF ANEST

不存在和存在 Anest 时五聚体离子通道的 X 射线结构

• 批准号: 8170297
• 负责人: PEI TANG
• 金额: $ 0.03万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2011-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8170297
• 关键词: Affinity; Anesthetics; Binding; Binding Sites; Breathing; Bromides; Cations; Chloride Ion; Chlorides; Complex; Computer Retrieval of Information on Scientific Projects Database; Data; Dose; Family; Funding; Gated Ion Channel; General Anesthesia; General anesthetic drugs; Grant; Halothane; Homologous Gene; Institution; Intravenous Anesthetics; Ion Channel; Ketamine; Ligands; Molecular; Proteins; Research; Research Personnel; Resolution; Resources; Source; Structure; Sulfur; Thiopental; United States National Institutes of Health; base; beamline; molecular size; novel; protein function

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The molecular mechanism of general anesthesia remains an enigma. Although a superfamily of pentameric ligand gated ion channels (pLGICs) has been identified as putative targets of general anesthetics, the lack of high-resolution structures of these pLGICs hinders the understanding where anesthetic binding sites are in these proteins and how anesthetic bindings impact on protein functions. An exciting platform for getting the answers has recently emerged as the x-ray structures of two bacterial homologs to the pLGIC family, GLIC and ELIC, were solved with resolutions of 2.9 and 3.3 ¿. As cation channels, GLIC and ELIC can be crystallized in the open- and close-channel states, respectively. Similar to mammalian pLGIC cation channels, cation conductance of GLIC could be inhibited by a variety of anesthetics at subclinical doses. Here we propose to co-crystallize GLIC and ELIC with general anesthetics. No anesthetic has been crystallized with any pLGICs in the past. The x-ray structures of GLIC- and ELIC-anesthetics complexes will provide novel structural basis to explain the functional impact of general anesthetics to the pLGICs. Inhaled anesthetic halothane and intravenous anesthetics thiopental and ketamine are chosen for the study. We have crystallized GLIC and ELIC in the absence and presence of anesthetics. Our preliminary data show that the resolutions up to 2.7 ¿ and 2.9 ¿ have been achieved for GLIC in the presence of thiopental and ketamine. Despite tangible protein resolutions, to define precise anesthetic binding sites in the proteins remains challenging because of small anesthetic molecular sizes and relatively low anesthetic binding affinities (~100 ?M). Thus, we request to use specific beamlines that provide high sensitivity and resolution to bromide, sulfur, and possibly chloride atoms. These atoms are contained in the selected anesthetics and can serve as marks for anesthetics in the proteinanesthetic complexes.

该副本是使用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这是调查员的机构。 全身麻醉的分子机制仍然是一种谜。尽管已将五聚体配件离子通道（PLGICS）的超家族确定为一般麻醉剂的推定靶标，但这些PLGIC的缺乏高分辨率结构阻碍了这些蛋白质中麻醉结合位点在这些蛋白质中以及麻醉对蛋白质对蛋白质的影响。最近出现了一个令人兴奋的答案平台，它是针对PLGIC家族的两个细菌同源物的X射线结构，即Glic和Elic，用2.9和3.3的溶液解决了。作为阳离子通道，可以在开放式和近频道状态下结晶的胶质和细胞。与哺乳动物的PLGIC阳离子通道类似，在亚临床剂量下，各种麻醉药可以抑制胶质的阳离子电导。在这里，我们建议与全身麻醉剂共结晶。过去，没有任何麻醉剂与任何PLGIC都结晶。胶质和精美 - 动作材料复合物的X射线结构将提供新的结构基础，以解释一般麻醉药对PLGICS的功能影响。吸入麻醉晕和静脉麻醉药硫代和氯胺酮用于研究。在不存在和存在麻醉药的情况下，我们已经结晶的胶质和精致。我们的初步数据表明，在硫胺和氯胺酮的存在下，胶质达到了高达2.7€和2.9。尽管有切实的蛋白质分辨率，但要定义蛋白质中精确的麻醉结合位点，由于小麻醉分子大小和相对较低的麻醉结合亲和力（〜100？m）仍然受到挑战。这是我们要求使用对溴化物，硫和可能的氯化原子的高灵敏度和分辨率的特定光束线。这些原子包含在选定的麻醉剂中，可以用作蛋白质消除复合物麻醉剂的标记。