Biochemistry is a field of study for students to build an interdisciplinary knowledge in the natural sciences. A Bachelor of Science in chemistry with an emphasis in biochemistry can lead to jobs in research, medicine, biotech and more. Here at CU, there are two projects that students can pursue.
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Protein folding and unfolding is often modeled as an equilibrium between two extreme states, folded and unfolded (F🡨🡪U). Differential Scanning Calorimetry (DSC) is a method by which heat flow between two samples is measured as a function of temperature increasing or decreasing. Proteins thermally denature, and thus transition between F🡨🡪U, at a specific temperature known as the melting temperature (TM). A DSC can measure protein unfolding by monitoring the difference in the heat flow between a protein sample dissolved in a buffer and a reference buffer. Heme binding proteins in pathogenic organisms like Listeria monocytogenes are virulence factors when causing infections. In order to investigate the thermal stability of these proteins, we plan to test two heme binding proteins from L. monocytogenes using a DSC-25. Further studies will elucidate the energy of heme (ligand) binding to the protein as well as investigating small molecules (drug) effects on protein stability.
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The F1Fo ATP synthase in E. coli is an essential enzyme complex that is responsible for most of the chemical energy inside the cell. The F1Fo ATP synthase can be targeted by antibiotics since bacteria need F1Fo ATP synthase to grow efficiently. High resolution models exist for all proteins in the complex except subunit a. Subunit a is the essential proton (H+) channel that couples the exergonic transport of H+ to ATP synthesis in the F1 portion. Recently, low resolution structural evidence has provided the best picture of the H+ channel in subunit a. However, previous biochemical data predicted a radically different channel structure compared to the new low resolution model. The structure of subunit a has a direct impact on its function as a proton channel and thus on the overall function of F1Fo ATP synthase. In this study we plan to compare and contrast the models by using Cys-Cys crosslinking to verify or refute predictions made by either model. Successful intra-subunit a crosslinks will be further investigated for their effect on function using a coupled fluorescent assay.
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