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Metals in Biology: From Metallotherapeutics to Cofactor Assembly and Trafficking

Fidai, Insiya N
http://orcid.org/0000-0002-2498-2411
http://rave.ohiolink.edu/etdc/view?acc_num=osu1471452195

Abstract Details

2016, Doctor of Philosophy, Ohio State University, Biophysics.
Metals are ubiquitous in nature and they perform vital functions in the cell that cannot otherwise be achieved, and therefore, biological systems utilize metals for diverse functions. This research focuses on two different areas – metals in medicine and metals in cofactor assembly and trafficking. Mankind has utilized metals in medicine dating back to 3000 BC. Discovery of cisplatin and ruthenium-based compounds laid the foundation for metallotherapeutics, making use of the unique properties of metals which can be superior to traditional drugs. One such approach is the development of catalytic metallodrugs which essentially recognize the therapeutic target like traditional drugs but in addition act like an enzyme, irreversibly modifying the target. This enables metallodrugs to be utilized with much lower dosage, act at sub-stoichiometric ratios, and also lead to lower side effects. This approach has been applied to two different targets; protein sortase A from Staphylococcus aureus and RNA from Hepatitis C virus. Catalytic metallopeptides that target the membrane-associated sortase A transpeptidase were developed and evaluated as irreversible inactivators of sortaseA, by modifying the active site residues Cys-184 to cysteine sulfonic acid and oxidation of the Arg-197 residue, which was supported by LC/MS-MS, NMR and other biochemical experiments. Another therapeutic target, Hepatitis C IRES RNA that was also investigated. Initial reports have demonstrated the activity of the Cu-GGHYrFK, copper peptide complex, in recognition of stem-loop IIb of the HCV IRES RNA. This research focuses on the structural and computational analysis of the stem-loop IIb of the HCV IRES RNA in complex with Cu-GGHYrFK and the all D-configuration of this peptide and further comparing the binding sites with the reactivity sites for these metallopeptides. Iron-sulfur clusters (Fe/S) are one of the most ancient, ubiquitous and versatile classes of metal cofactors found in nature. Proteins that contain Fe/S clusters constitute one of the largest classes of proteins, with varied functions that include electron transport, regulation of gene expression, substrate binding and activation, and radical generation. Some of the Fe/S cluster-binding proteins have been characterized, but mechanisms of cluster biosynthesis, transport, and exchange are not well established. The focus of this study is to identify the most physiologically relevant trafficking pathways based on quantitative evaluation of cluster transfer between pairs of putative protein partners using spectroscopic and kinetic assays. A detailed study of the kinetic transfer rates of the [2Fe-2S] cluster proteins, which include scaffold proteins, glutaredoxins, cluster carriers proteins, and the final target proteins was performed. Based on our results, I have developed two working models, one for cytosolic and another one for mitochondrial Fe/S cluster biosynthesis and trafficking, providing mechanistic and kinetic details of cluster transfer steps. In addition to mapping Fe/S cluster uptake and transfer pathways, the structural, functional and biochemical characteristics of Synechocystis NifU, a [2Fe-2S] cluster binding protein was also investigated. Collectively, the results from this research project provide novel insights in to Fe/S cluster biogenesis and trafficking and its role in several Fe/S cluster related diseases.
James Cowan (Advisor)
Richard Swenson (Committee Member)
Dalbey Ross (Committee Member)
Bell Charles (Committee Member)
322 p.
Biophysics

Recommended Citations

Citations

  • Fidai, I. N. (2016). Metals in Biology: From Metallotherapeutics to Cofactor Assembly and Trafficking [Doctoral dissertation, Ohio State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=osu1471452195

    APA Style (7th edition)

  • Fidai, Insiya. Metals in Biology: From Metallotherapeutics to Cofactor Assembly and Trafficking. 2016. Ohio State University, Doctoral dissertation. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=osu1471452195.

    MLA Style (8th edition)

  • Fidai, Insiya. "Metals in Biology: From Metallotherapeutics to Cofactor Assembly and Trafficking." Doctoral dissertation, Ohio State University, 2016. http://rave.ohiolink.edu/etdc/view?acc_num=osu1471452195

    Chicago Manual of Style (17th edition)

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This open access ETD is published by The Ohio State University and OhioLINK.