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

描述(由申请人提供)：K01奖将使Harry Gill博士从一名目前受指导的助理教授转变为一名独立的科学研究员，专注于了解肾脏NBCe1-A的Na/HCO3共转运机制。吉尔博士的背景是分子生物学、生物化学和结晶学。然而，由于必要的功能分析是生理学的，因此将吉尔博士整合到他的导师沃尔特·博伦教授的生理学实验室是至关重要的。一个由来自三个机构的资深调查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)在细胞水平上开发新的实验(电生理)技术；(B)在全动物水平上为新的实验工作做准备；(C)教授生理学课程；(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Mentor一起参加Mentor以生理为导向的科学会议；(G)启动由Mentor生成的网络；(H)进行(有限的)教学以加强培训；(I)在实验室指导本科生；以及(J)在助学金撰写方面的培训。(D)项将为吉尔博士提供初步数据，并为他的第一次R01申请做好准备。该研究计划的具体目的是：(1)利用X射线结晶学来阐明胞质N末端(NT)和人类R298S突变在NBCE1-A功能中的作用。(2)培养细胞生理学家的电生理技能，利用由内而外的巨型贴片首次测定NBCe1-A对胞质[Na]、[HCO3]和pH的依赖性。(3)探讨穿透NBCe1胞质NT内部的管道的功能意义，并为下一代携带关键NT突变的敲入小鼠制备靶向载体。拟议的实验工作-从原子到细胞/分子水平-将为细胞质NT在更广泛的SLC4家族成员的功能中的作用提供重要的新见解。K01培训计划将培养出一位全面发展的学术科学家，他将在提交自己的R01补助金方面具有很强的竞争力。

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.