How can we transform the undergraduate laboratory curriculum to resemble the research laboratory and beyond?  

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This is a question that has been on my mind constantly since I started my PhD program. I was an undergraduate who did well in lecture and enjoyed laboratory research, but was intimidated by the laboratory portion of my courses. After graduation, despite my extensive undergraduate research experience, I struggled to adapt to graduate school during my first and second years. With practice, patience and a lot of support, I succeeded! But I became inspired to develop inquiry-based laboratory experiences for undergraduate students that encouraged the development of creative inquiry. 

 

My ultimate goal as an educator is to develop a curriculum that branches the world of the undergraduate classroom with the world after graduation. I achieve this by combining my broad and extensive protein biochemistry research expertise with my other passions--education and the arts--to develop a wholistic approach to chemistry that reflects the spirit of creative inquiry and critical thinking that is the hallmark of a liberal arts education. I incorporate the stories of real-world scientists, historical events and social justice issues with to foster an inclusive and anti-racist classroom. Below, I detail my experiences to date in laboratory development, and my research experience as a biochemist. 

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Classroom and Laboratory Development

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Dyeing to Learn Chemistry: At-Home Dyeing With In-House Kits

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At Davidson College, I had the opportunity to develop my own non-natural science majors' course on the chemistry of Fibers and Dyes, called Dyeing to Learn Chemistry. The best part was designing a laboratory that exposed students to different types of dyes as a visualization of the chemistry occurring at the molecular level. When my classes for Fall 2020 and Spring 2021 were remote, however, I pivoted, developing laboratory kits that could be used in the common area, dorm room, or at-home kitchen. With these kits, students explored the effects of time and concentration on hibiscus and butterfly pea tea intensity, and the effects of three different metal mordants on cochineal dye. 

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An in-depth analysis of these experiments, as well as the virtual experiments and experiments performed in a related course for upper-level chemistry majors designed by my colleague, Angela G. King PhD, will soon be available in our ACS Symposium Series Chapter entitled "Teaching Undergraduate Chemistry Through Fibers and Dyes."

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Want to see some dye chemistry in action? Check out these instagram stories on pH and madder, effects of time and concentration, the chemistry of tie dye, and preparing a natural indigo dye vat. Note: you need an Instagram account to access these links. 

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How I used Instagram stories to communicate laboratory content to remote students during the COVID-19 pandemic in hte Spring 2020 semester is highlighted in a joint publication with my colleagues in the chemistry department at Davidson College.

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Publications

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King, A.G. and Gorensek-Benitez, A. H. “Teaching undergraduate chemistry through fibers and dyes.” 2021. In Chemistry in

Context: Teaching chemistry concepts in the context art and archeology; Braun, K. and Labby, K., Eds.; ACS Symposium

Series. Invited chapter. In Press.

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Anstey, M. R., Carroll, F. A., Gorensek-Benitez, A. H., Hauser, C. D., Key, H. M., Myers, J. L., Stevens, E P., Striplin, D.,

Holck, *H. W., *Montero-Lopez, L. M. and Snyder, N. L. “#DavidsonTrue: Transitioning to Remote Teaching while

Maintaining our Values as a Liberal Arts College During the COVID-19 Pandemic at Davidson College.” 2020. Journal of

Chemical Education. 97:2800-2805.

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Bringing Research to Lab: Using the Agilent Seahorse to Manipulate Glycolysis in Real-Time

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As a postdoctoral teacher-scholar fellow at Wake Forest University, I collaborated with Dr. Bruce King to develop an undergraduate curriculum that integrated cellular metabolism in the teaching laboratory. Using the Agilent Seahorse Extracellular Flux Analyzer, I developed a laboratory module that analyzed the cellular response to inhibition of lactate dehydrogenase, the enzyme purified and characterized by students in lab over the course of the semester. Through this laboratory, students were given the opportunity to visualize, at the cellular level, a response to inhibition of a single enzyme.

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You can read more about mine and Bruce's work here. 

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Biochemistry Research Experience 

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Since learning about protein structure as an undergraduate student, I have endeavored to characterize the thermodynamics and kinetics of various proteins--for folding, function, and fibrillation. Most of my research has centered on understanding how the crowded cellular interior--which contains upwards of 300 g/L of proteins for an E. coli cell--affects how proteins work. Since most bench biochemistry occurs in dilute solution with less than 10 g/L of other macromolecules, this research can inform how we interpret and apply dilute solution studies of protein folding and function to the cellular environment. 

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Elucidating the Effects of Macromolecular Crowding on Alpha-Synuclein Fibrillation

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This research was performed in collaboration with Jeff Myers at Davidson College and four undergraduate researchers. While my research to this point had been dedicated to characterizing protein function, this research project is dedicated to understanding protein dysfunction--or how macromolecular crowding affects the aberrant fibrillation of alpha-synuclein, the protein whose fibrillation and plaque formation is a hallmark of Parkinson's Disease. With undergraduate students, we explored the effects of concentrations of sugar-based polymers and their constituent monomers on fibrillation. 

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Measuring the Pre-Steady State Kinetics of Methionine tRNA Synthetase

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This research was performed during my post-doctoral fellowship in Rebecca Alexander's laboratory at Wake Forest University. I used a KinTek stopped-flow instrument to characterize the pre-steady state kinetics of three different enzymes: the engineered monomeric E. coli methionine tRNA synthetase (MetRS), the native dimeric E. coli MetRS, and another dimeric MetRS from S. cerevisiae. While performing this research, I mentored three undergraduate research students. 

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Determining How Macromolecular Crowding Modulates Protein Folding

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This area of research formed the bulk of my graduate research in Gary Pielak's lab at UNC Chapel Hill. In this work, I sought to understand how macromolecular crowding influences the protein folding kinetics of the n-terminal SH3 domain of the Drosophila protein drK (SH3). SH3 is a metastable protein, which exists in close to equal amounts of the folded state and unfolded ensemble at equilibrium. The stability of the protein, which was fluorinated at its single tryptophan residue, combined with the slow chemical exchange, allows us to observe both states, and the kinetics of their interconversion, using 19F-detected Nuclear Magnetic Resonance Spectroscopy. 

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During my PhD research I developed a protocol to purify this protein. I measured the folding and unfolding rate constants in the presence synthetic polymers and their monomers, as well as urea and the protein lysozyme. Making these measurements at several temperatures allowed me to determine the temperature dependence, and therefore the the activation free energy, enthalpy and entropy of folding and unfolding. I mentored one undergraduate research student in this work. 

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Publications

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Gorensek-Benitez, A. H., Smith, A. E., Stadmiller, S. S., Perez Goncalves, G. M. and Pielak, G. J. “Cosolutes, crowding and

protein folding kinetics.” Journal of Physical Chemistry B. 121: 6527-6537

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Stadmiller, S. S., Gorensek-Benitez, A.H., Guseman, A. J. and Pielak, G. J. “Osmotic-shock induced protein destabilization

in living cells and its reversal by glycine betaine.” Journal of Molecular Biology. 429:1155-1161.

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Smith, A.E., Zhou, L.Z.,* Gorensek, A.H., Senske, M. and Pielak, G.J. 2016. “In-cell thermodynamics and a new role for

protein surfaces.” Proceedings of the National Academy of Sciences USA. 113: 1725-1730

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Studying the Effects of Small and Large Cosolutes on Enzyme Activity

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This research project was drive by the efforts of two undergraduate students. The goal of this project was to understand how size, concentration and, ultimately, the conformation, of synthetic polymers and their constituent monomers affected the activity of the enzyme, E. coli dihydrofolate reductase, and enzyme involved in purine metabolism that is the target of the anti-cancer therapeutic, methotrexate. Also in this work, we compiled the results of many studies that explored the effects of macromolecular crowding on enzyme kinetics, with the ultimate determination that crowding effects are idiosyncratic, vary from protein to protein, and are more difficult to predict than we had originally anticipated. 

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Publications

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Acosta, L.C., Perez Goncalves, G. M., Pielak, G. J. and Gorensek-Benitez, A. H. “Large cosolutes, small cosolutes and

dihydrofolate reductase activity.” Protein Science. 26:2417-2425.