Cellular lipid transport determined with multifunctional lipid derivatives

项目摘要 在我们目前的R01项目中，我们使用多功能脂质衍生物来鉴定脂质结合蛋白，并 确定完整细胞内的脂质位置。后者我们用来研究细胞膜上的脂质运输。 系统连接到另一个系统。明确地说，我们研究了新的假定的脂转运蛋白及其在细胞信号转导中的作用。 和功能。我们鉴定了两种蛋白质，胞质MPP6和质膜相关的ATP11A。 暑期学生将准备MPP6的单一残基突变体，并分离突变蛋白。这位学生 将调查突变体对使用脂条的脂类结合的影响以及脂类运输的变化 通过CRISP/Cas9技术用突变品种替换内源MPP6。时间允许的话 学生还将准备ATP11A的第一个GFP融合蛋白。阿米莉亚·科塔玛蒂女士的候选人 在克隆技术和组织培养方面有经验，熟悉我们的荧光激光扫描共聚焦 显微镜。这名学生将由经验丰富的技术员安德鲁·古斯塔夫森先生指导。这位学生 将参加我们的小组会议、实验室活动以及暑期本科生研究 化学生理和生物化学系计划(SURP)。

Project Summary In our current R01 project, we use multifunctional lipid derivatives to identify lipid binding proteins and to determine lipid location inside intact cells. The latter we use to study lipid transport from one cellular membrane system to another. Explicitly, we investigate new putative lipid transport proteins and their role in cell signalling and function. We identified two proteins, the cytosolic MPP6 and the plasma membrane-associated ATP11A. The summer student will prepare single residue mutants of MPP6 and isolate the mutant proteins. The student will investigate the influence of the mutants on lipid binding by using lipid strips and changes in lipid transport by replacing the endogenous MPP6 by the mutant variety via the CRISP/CAS9 technique. Time permitting the student will also prepare the first GFP fusion proteins of ATP11A. The candidate Ms. Amelia Kotamarti has experience in cloning techniques and tissue culture and is familiar with our fluorescent laser-scanning confocal microscope. The student will be supervised by an experienced technician, Mr. Andrew Gustafson. The student will participate in our group meetings, and lab activities as well as in the Summer Undergraduate Research Program (SURP) of the Department of Chemical Physiology and Biochemistry.