This CAREER award from the Biomaterials Program in the Division of Materials Research to Boise State University is to study biophysical properties of a specific pore-forming protein isolated from earthworms. These pore forming proteins will be studied for their unique control over the transport of relevant molecules. The scientific broader impact of this project is the development of efficient methodologies for molecular identification, single-molecule analysis, design of nano-containers for controlled delivery of chemicals and drugs, and cell loading through temporary membrane permeabilization. The research activities will be incorporated into interdisciplinary teaching and mentoring practices for high-school, undergraduate and graduate students. Specific student recruitment approaches are in place to ensure participation of women, underrepresented, and economically disadvantaged students in research and education. The investigator will be actively engaged in outreach activities at local schools and educational centers, will develop the students' ability to effectively share their interest in science, and to energize young students to pursue STEM careers and to cultivate a greater understanding and appreciation of the merits of scientific research.
The research objective of this CAREER award is to investigate the ability of the pore-forming protein lysenin isolated from earthworms to function as either a nano-sensing device or as nano-valve actuated by physical or chemical stimuli. The large size of the lysenin pore opening will enable use of the resistive pulse technique for stochastic sensing, and single-molecule detection and characterization. Reversible ligand-gating induced by multivalent cations is exploited to achieve exquisite control over the transport of molecules across artificial lipid membranes that are being prepared. Specific aims of the project are; 1) to investigate the transport of macromolecules through lysenin channels inserted into artificial membranes; 2) to initiate and control the transport through artificial lipid membranes with lysenin channels; and 3) to exploit lysenin voltage regulation for reversible control over cell membrane permeability and to develop an understanding of the conducting state of lysenin channels with external electric fields. These studies would advance and significantly transform the current techniques and methodologies employed for fast and reliable molecular sizing, single-molecule analysis and sequencing, design of nano-containers for controlled delivery of chemicals and drugs, and cell loading through temporary membrane permeabilization. The project includes a detailed and carefully tailored plan for recruiting, mentoring and teaching of students, and outreach activities. Undergraduate, graduate, and high-school students will actively participate in this cutting-edge interdisciplinary research utilizing electrophysiology, fluorescence spectroscopy, and molecular biology techniques. The outreach activities developed with the participant students are expected to cultivate a greater understanding of the merits of scientific research to the public and mobilize young students to pursue careers in STEM areas.
