MRI: Track 1 Acquisition of an FPLC System for Research and Training at Winona State University

Winona State University, Winona MN

Investigators

Abstract

This award is funded by the Major Research Instrumentation Program and the Chemistry Research Instrumentation Program. Associate Professor Emily F. Ruff from Winona State University, with a team of three faculty from the Chemistry department, is acquiring a fast protein liquid chromatography (FPLC) instrument and accompanying software and accessories for research and training. FPLC is a technique used to purify and analyze proteins with high reproducibility, yield, and speed. Product proteins can be used in high-quality downstream biochemistry and biophysics experiments. This acquisition will modernize the biochemistry and molecular biology infrastructure in our department, and it will provide training opportunities to prepare students for work in research and industry. This award aims to enhance several projects in research and teaching. FPLC instruments can be used to run pre-packed affinity, ion exchange, and size exclusion columns for protein purification and analysis, with automated control of buffer gradient and flow rate and constant monitoring of pressure, conductivity, and UV-Vis absorbance. FPLC will be used to purify protein kinases such as human vaccinia-related kinase 1 (VRK1) from a bacterial expression system. The structure and stability of these variants will be compared using techniques like circular dichroism and differential scanning fluorimetry, which may shed light on their role in disease. The FPLC instrument will also be also used for protein purification and analytical gel filtration of Drosophila melanogaster neuroblast complexes and proteins. This will help to elucidate the roles of proteins like Inscuteable, Pins, and Bazooka in controlling asymmetric cell division. Variants of the enzyme polyethylene terephthalate hydrolase (PETase) will also be purified for development of course-based undergraduate research experiences (CUREs), engaging upper-level and introductory chemistry students in thinking about the world’s plastic problem. Use of FPLC will also be explored for a teaching laboratory exercise examining how post-translational modifications of lysozyme affect amyloid formation and misfolding. Members of our team have been working on related projects using gravity flow chromatography, but using FPLC techniques will strengthen and professionalize these projects and improve the quality of collected data. Undergraduate students will be involved at all levels of this work. They will be trained in high-level techniques used in industry and research, and some may even become co-authors on peer-reviewed publications. Use of this instrumentation may also enable collaborations with local partners and recruitment and retention of new faculty and students. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.