Date of Graduation
5-2026
Document Type
Thesis
Degree Name
Bachelor of Science in Biology
Degree Level
Undergraduate
Department
Chemistry & Biochemistry
Advisor/Mentor
Dr. Suresh Kumar Thallapuranam
Committee Member
Dr. Adnan Alrubaye
Second Committee Member
Dr. Yuchun Du
Abstract
Fibroblast Growth Factors (FGFs) are critical signaling molecules that regulate essential processes such as cell proliferation and tissue repair. While FGF-1 is a "universal" growth factor, its therapeutic utility is limited by poor thermal stability and a short half-life. An engineered variant, Super FGF-1 (sFGF1), offers hyperstability but demonstrates reduced heparin-binding affinity. This project explores the design of a novel sFGF1-FGF10 chimera by introducing three Nterminal mutations - C10K, S11K, and N12G - to restore heparin affinity while maintaining stability. Experimental results confirmed successful bacterial transformation and soluble protein expression. Fluorescence spectroscopy and Circular Dichroism (CD) spectroscopy revealed that these mutations triggered a structural transition from the native beta-sheet framework to an alpha-helix secondary structure, accompanied by a peak intensity shift to 330 nm. Furthermore, the chimera elutes at 300 mM and 500 mM NaCl, confirming restored heparin affinity. Despite a less compact hydrophobic core as indicated by 8-anilino-1-naphthalenesulfonic acid (ANS) binding assays, the variant maintains high thermal stability with a Tm of 67°C. These findings provide a stable foundation for the development of advanced wound-healing therapeutics.
Keywords
Fibroblast; Protein; Characterization; FGF-1; sFGF1; Heparin
Citation
Anandaraj, K. (2026). Design and Construction of 3 Point Mutations (C10K/S11K/N12G) Towards the Super FGF1- FGF10 Chimera. Chemistry & Biochemistry Undergraduate Honors Theses Retrieved from https://scholarworks.uark.edu/chbcuht/74
