The protein actin is integral to movement, cell adhesion, and cytoskeleton support within the human body. Actin is the most prominent protein within cells, and it participates in more protein-protein interactions than any other known protein; one such relationship involves palladin. Palladin is comprised of five immunoglobulin-like domains (Ig), each connected via an unstructured linker region. Previous research has proven that the Ig3 domain is the minimal actin-binding domain, meaning it is the only domain required to facilitate binding between palladin and actin; however, binding affinity is significantly increased when the Ig3-4 linker domain is present. To examine the effects of this Ig3-4 linker on overall actin binding, the Beck lab introduced several mutations into the regions. When all the arginines within Ig3-4 were converted to alanine, the binding ability of actin was completely disrupted. Our current research seeks to determine how mutations within the Ig3-4 linker region will affect the overall structure and stability of palladin. Our study was conducted with both wild-type and mutant arginine to alanine linker proteins. These proteins were purified from pellets and then examined using circular dichroism (CD) spectroscopy. Our proteins were placed under wavelength scans of 185-250 nanometers, as well as thermal denaturation conditions, both of which revealed slight differences between the curves of wild-type and mutant palladin. This information was then analyzed using DichroWeb software, which uses input data to create charts detailing folding, structure, reconstructed data curves, and suspected wavelength impacts. A preliminary review of this data indicates that the folding of mutated Ig3-4 linker contains more random coils and alpha helix structures while forming significantly fewer beta-sheet folds. Future research will involve protein comparisons of these structures using the PCDDB database, a public site where the structure of wild-type and mutated Ig3-4 linker regions will be published.