The most abundant protein in eukaryotic cells is actin, which is highly conserved and has multiple protein interactions. These interactions enable actin to perform various functions such as muscle contractions, cell division, cell migration, and cell adhesion. Myopalladin (MYPN) is one of the many actin-binding proteins that interact with actin through protein-protein interactions. It is a Z-disc protein made of immunoglobulin-like (Ig) domains that interact with actin to regulate the structure and function of cardiac muscle. Myopalladin affects dynamics and cardiac muscle growth while maintaining Z-disc integrity during aging and exercise. Analyzing the interaction of MYPN and actin will help further the understanding of the role of myopalladin in maintaining the Z-disc structure and function of cardiac muscle. Since the surface of F-actin is acidic and the surface of Ig3 MYPN is basic, we hypothesize that electrostatic interactions drive complex formation, and mutations of basic residues in MYPN will affect this interaction significantly. Prior research in the Beck lab used crosslinking-mass spectrometry (XL-MS) data to predict a model structure of the crosslinked proteins, and these point mutations will provide further evidence that these basic regions of myopalladin are essential for interactions with F-actin. Based on previous data, there is successful crosslinking of myopalladin’s Ig3 domain with F-actin using a DMTMM crosslinker. This study aims to investigate the effect of point mutations of basic residues identified in the XL-MS-based model of this complex between F-actin and the minimal actin-binding domains of myopalladin. The experiment entails using site-directed mutagenesis and polymerase chain reaction to mutate the K43 base sequence in the MYPN Ig3 domain from lysine to alanine. After this, protein purification and actin co-sedimentation assays will be used to measure actin binding to mutated residues. This study could provide insight into future research about the potential of these mutations to cause phenotypic changes in z-disc structure and function in the sarcomere region of cardiac muscle.