| | Category | BI | L06 | Identifying Biophysical Properties of Iron Sulfur Cluster Assembly |
| | Proteins |
| | Abstract | Iron-sulfur proteins are important in cellular processes such as iron |
| | homeostasis and are also an integral part of our neurological functions. |
| | Iron sulfur clusters are found in all living organisms and serve as catalysts |
| | in chemical reactions, electron carriers in redox reactions, regulators of |
| | iron homeostasis, and stabilizers of protein structure. The aim of this |
| | project is to investigate biophysical characteristics and interactions of Fe- |
| | S proteins involved in the ISC assembly. The main focus lies on the |
| | essential role of Isu, the scaffold protein in the mitochondrial ISC |
| | construction, which combines the iron protein Frataxin and the sulfur |
| | protein NfsI. In this study, a mutant of the iron protein frataxin was |
| | isolated, purified, and studied to determine its binding ability with respect to |
| | Isu. Protein growth, isolation, and purification through Fast Protein Liquid |
| | Chromatography and gel electrophoresis were used in order to isolate Isu |
| | protein as well as Yeast and Drosophila Frataxin, the protein responsible |
| | for transporting iron to the binding site. The isolated and purified proteins |
| | were often concentrated through lyophilization, or freeze drying. After |
| | protein purification and concentration, activity assays were performed to |
| | test and confirm the role of Isu and Yeast Frataxin by comparing the |
| | amount of Fe-S clusters formed. In additional research, Isothermal Titration |
| | Calorimetry was also performed in order to learn more about the |
| | biophysical structure and binding affinity of each protein. The results of |
| | these tests confirm Isu's role as an essential protein in Fe-S cluster |
| | assembly. It also suggests that the Yeast Frataxin Mutant studied had only |
| | one iron binding site but still allowed for increased formation of iron sulfur |
| | clusters. These results can be used to better understand this constant and |
| | vital process of ISC assembly, which is thought to be among the earliest |
| | catalytic processes in the evolution of biomolecules. |
| | Bibliography | R. Lill and U. Muhlenhoff. Maturation of Iron-Sulfur Proteins in Eukaryotes: |
| | Mechanisms, Connected Processes, and Diseases. Annual Review |
| | Biochem. 2008. |