Doctor of Philosophy, The Ohio State University, 2017, Biophysics
Metalloproteins are essential to life, with roles in cellular signaling, metabolism, gas cycles, and gene regulation, among others. One group of metalloproteins is the ferritin-like superfamily, which contains a wide variety of members ranging from iron-trafficking proteins (ferritin) to enzymes that can perform 2-electron chemistry on hydrocarbons (bacterial multicomponent monooxygenases or BMMs). Ribonucleotide reductases (RNRs), one of the members of the ferritin-like superfamily, are enzymes that catalyze the only known de novo reduction of ribonucleotides to deoxyribonucleotides and are found is nearly every organism on earth. Recently, a novel protein was discovered in Mycobacterium tuberculosis called R2lox, because of its sequence similarity to the R2 subunit of RNR, whose function remains unknown. However, it does not possess any RNR activity and is more similar in active site structure and proposed function to the BMMs. This protein is very important from a basic science standpoint as well as a health science standpoint because of its unique metal-binding capabilities and because it is upregulated in the virulent H37rv strain of M. tuberculosis. To study the active site structure and oxygen activation reaction of this protein, a custom resonance Raman spectroscopic system was designed and built. This system is very versatile in that it can access many different wavelengths with a single laser, which is useful for resonance Raman, which relies on tuning the Raman excitation beam to an electronic absorption to enhance the scattering efficiency of Raman light. In addition to R2lox, this custom resonance Raman system has been used on numerous bioinorganic molecules to great effect. This work describes the design and construction of the custom resonance Raman system and its use in studying R2lox and other bioinorganic systems. Additionally, experiments were performed to lay the groundwork for studying electron transfer in R2lox via ruthenium modification.
Committee: Hannah Shafaat PhD (Advisor); James Cowan PhD (Committee Member); Terry Gustafson PhD (Committee Member); Marcos Sotomayor PhD (Committee Member)
Subjects: Chemistry