Cellular lipid transport determined with multifunctional lipid derivatives

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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.