Structure-function studies of the dimerization of the N-terminal domain of NBCe1-

NBCe1- N 端结构域二聚化的结构功能研究

• 批准号: 8417670
• 负责人: Harry Singh Gill
• 金额: $ 14.03万
• 依托单位: GEORGE WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-21 至 2014-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8417670
• 关键词: Acid-Base Equilibrium; Acidosis; Acids; Affect; Air; Animals; Area; Bicarbonates; Binding; Biochemistry; Biological Assay; Blood; Boron; Boxing; Cartoons; Cell physiology; Cells; Charge; Computer Simulation; Coupled; Crystallization; Crystallography; Cytosol; Data; Dependence; Dependency; Dimerization; Educational process of instructing; Electrolyte Disorder; Electrophysiology (science); Eye; Family; Family member; Figs - dietary; Future; Future Generations; Genes; Gills; Grant; Health; Helix-Turn-Helix Motifs; Hereditary Disease; Human; Human Genetics; Institution; Ions; Journals; Kidney; Knock-in Mouse; Lead; Learning; Liquid substance; Measures; Membrane; Membrane Potentials; Membrane Proteins; Mental Retardation; Mentored Research Scientist Development Award; Mentors; Mining; Modeling; Molecular; Molecular Biology; Mus; Mutate; Mutation; N-terminal; Oranges; Patients; Physiological; Physiology; Play; Preparation; Process; Property; Proteins; Research; Research Personnel; Resolution; Role; Scientist; Side; Solvents; Sorting - Cell Movement; Structure; Surface; Techniques; Testing; Time; Tissues; Training; Training Activity; Training Programs; Transmembrane Domain; Urine; Work; Writing; X-Ray Crystallography; arm; base; basolateral membrane; dimer; improved; insight; interest; kidney cell; kidney epithelial cell; meetings; member; monomer; mutant; novel; novel therapeutics; physical model; prevent; professor; skills; solute; structural biology; undergraduate student; vector

DESCRIPTION (provided by applicant): The K01 award will transition Dr. Harry Gill from a currently mentored Assistant Professor to an independent scientific investigator, focusing on understanding the mechanism of Na/HCO3 cotransport by renal NBCe1-A. Dr. Gill's background is in molecular biology, biochemistry, and crystallography. However, because the necessary functional assays are physiological, it is critical that Dr. Gill be integrated into the Physiology Lab of his mentor, Professor Walter Boron. A mentoring committee consisting of senior investigators from three institutions will guide Dr. Gill's progress. The training activities include: (a) developing new experimental (electrophysiological) techniques at the cellular level; (b) preparing for new experimental work at the whole-animal level; (c) taking didactic physiology coursework; (d) performing original research; (e) participating in lab meetings, journal clubs, and seminars; (f) attending mentor at physiologically oriented scientific meetings with mentor; (g) initiating mentor-generated networking; (h) teaching (limited) to enhance training; (i) mentoring undergraduate students in the Lab; and (j) training in grant writing. Item (d) will provide Dr. Gill with preliminary data and otherwise prepare him for his first R01 application. The specific aims of the research plan are: (1) To use X-ray crystallography to illuminate the role of the cytosolic N terminus (Nt) and the human R298S mutation in the function of NBCe1-A. (2) To develop electrophysiological skills as a cell physiologist, and exploit the inside-out giant patch to measure for the first time the dependence of NBCe1-A on cytosol-side [Na], [HCO3], and pH. (3) To explore the functional significance of conduits that appear to weave through the interior of the cytosolic Nt of NBCe1, and to prepare a targeting vector for the future generation of a knock-in mouse carrying a key Nt mutation. The proposed experimental work-from the atomic to the cellular/molecular levels-will provide important new insights into the role of the cytosolic Nt in the function of members of the broader SLC4 family. The K01 training program will produce a well rounded academic scientist who will be highly competitive in submitting his own R01 grant.

描述（由申请人提供）：K 01奖将使Harry Gill博士从目前接受指导的助理教授转变为独立的科学研究者，专注于了解肾脏NBCe 1-A的Na/HCO 3共转运机制。Gill博士的背景是分子生物学、生物化学和晶体学。然而，由于必要的功能测定是生理性的，因此将Gill博士纳入他的导师Walter Boron教授的生理学实验室至关重要。一个由来自三个机构的高级研究人员组成的指导委员会将指导吉尔博士的进展。培训活动包括：（a）开发新的实验性在细胞水平上的（电生理学）技术;（B）为整个动物水平上的新实验工作做准备;（c）参加教学生理学课程;（d）进行原创研究;（e）参加实验室会议、期刊俱乐部和研讨会;（f）与导师一起参加面向生理学的科学会议;（g）启动导师生成的网络;（h）教学（有限）以加强培训;（i）在实验室指导本科生;以及（j）编写赠款的培训。项目（d）将为吉尔博士提供初步数据，并为他的第一次R 01申请做好准备。本研究计划的具体目标是：（1）利用X射线晶体学方法研究NBCe 1-A胞浆N端（Nt）和人R298 S突变在NBCe 1-A功能中的作用。(2)培养细胞生理学家的电生理学技能，并首次利用由内而外的巨大斑块来测量NBCe 1-A对胞质侧[Na]、[HCO 3]和pH的依赖性。（3）探索似乎穿过NBCe 1胞质Nt内部的管道的功能意义，并制备用于携带关键Nt突变的基因敲入小鼠的未来一代的靶向载体。拟议的实验工作-从原子到细胞/分子水平-将提供重要的新的见解胞质Nt在更广泛的SLC 4家族成员的功能中的作用。K 01培训计划将培养出一位全面发展的学术科学家，他在提交自己的R 01资助方面将具有高度竞争力。