All cells maintain ion gradients across the plasma membrane, which provides energy for multiple physiological processes. This gradient is established and maintained by the Na/K pump, a P-type 2 ATPase. Members of this protein class are present in all phyla and transport different metals with nearly identical mechanisms. Ion recognition and ion driven conformational changes are an essential part of this universal mechanism that gives singularity to each ATPase and are poorly understood. This project focuses on P-type IIC ATPases, proteins expressed by all animals, whose function is essential for adaptation to different salinities, digestion and cellular homeostasis. The research will use a combined approach of molecular biology, high-temporal resolution electrophysiology and fluorescence techniques to study the molecular mechanisms that allow recognition of the transported ions by P-type IIC ATPases and how ion transport is controlled by protein conformational changes. In particular, structural information from crystal structures of P-type IIC ATPases will be used to target changes in the regions that bind ions and in intracellular regions of the proteins, with functional affects monitored by voltage clamp techniques combined with fluorescence. The results from this project will provide critical insight and hopefully resolve prevailing models regarding the molecular mechanics of the P-type ATPase superfamily.
BROADER IMPACTS The broader impacts goal of this project is to inspire undergraduate students to pursue careers in fundamental basic sciences, and to develop the technical skills and attitudes towards discovery through scientific research. The PI's laboratory has a history of involving undergraduate and graduate students in scientific discovery, with trainees serving as co-authors on articles in high-impact journals, as well as presenting their findings at national scientific meetings. Several of trainees have pursued scientific careers. This project will involve undergraduate students from various programs and disciplines, including a focus on the recruitment of underrepresented minorities from institutional programs such as the Plains Bridges to Baccalaureate program. The results from this project will be disseminated by publications in high-impact journals, courses, invited lectures and presentations at high visibility national and international scientific meetings.
