CAREER: Biosensor Development for Probing Nanoscale Topology in Neurotransmission

University Of Denver, Denver CO

Investigators

Abstract

1452057 - Knowles Sensors that measure biological molecules are essential for medical diagnostics and drug discovery. These sensors often rely on the measurement of membrane proteins, which are targets for drug discovery. On the other hand, biomimetic systems provide simple platforms for incorporating membrane proteins into biosensing applications. In these applications, membrane proteins are able to sense their local environment, such as the chemical composition and membrane shape, and these factors affect protein function. Until now little attention has been given to the nanoscale structure of these sensors and how nanometer-sized features affect proteins. The main focus of this research is to design biosensors that can be used to identify how cells and biomolecules are affected by nanostructured materials. Specifically, proteins involved with the transmission of neuronal signals and the secretion of hormones will be characterized. The integration of research with education will take place through outreach to 8th-12th grade students in a summer engineering camp, training of local educators during a summer research program, and the design of novel courses at the interface of biochemistry and engineering. The engineering of materials that interface with biological systems increasingly requires an understanding of cellular and molecular responses to nanoscale topology. The focus of this CAREER Award is to design two biosensors that will be used to probe the relationship between nanostructured materials and protein function. The first sensor will mimic the intracellular plasma membrane with tunable regions of membrane curvature and chemical composition. The second will provide a template to introduce membrane curvature into live cells, where the molecular response to curvature can be assessed. Both sensors will be used to characterize the relationship between membrane shape and neurotransmission, a biological process that gives rise to extreme changes in membrane topology. Neurotransmission relies on the proper recruitment and function of SNARE proteins to facilitate the fusion of the tethered vesicle membrane with the plasma membrane. SNARE-mediated membrane fusion is essential for membrane repair, growth cone formation, axon extension, and synapse formation. Our work aspires to contribute to the field of neuroregeneration by identifying nanoscale features that drive membrane fusion. By using super-resolution fluorescence microscopy techniques, single particle tracking, and combined confocal fluorescence-atomic force microscopy, principles that govern protein sorting on nanostructured membranes will be identified. These principles will be useful in the future design of biosensors and materials that interface with cells. The integration of this research with education will take place through outreach to 8th-12th grade students in a summer engineering camp, training of local educators during a summer research program, and the design of novel courses at the interface of biochemistry and engineering.