Design of an FGF-1-FGF-2 Heterodimer Variant with Enhanced Stability and Cell Proliferation Activity
Date of Graduation
12-2023
Document Type
Thesis
Degree Name
Bachelor of Science in Chemistry
Degree Level
Undergraduate
Department
Chemistry & Biochemistry
Advisor/Mentor
Thallapuranam, Suresh
Committee Member/Reader
Aloia, Lindsey
Committee Member/Second Reader
Dong, Bin
Committee Member/Third Reader
Chapman, Kate
Abstract
The FGF-1 subfamily, composed of FGF-1 and FGF-2, assists in broad health-related processes such as cell proliferation and angiogenesis respectively.1,2 The subfamily shows promising signs as a therapeutic, however, the inherent thermal instability leads to a low half-life in vivo.3 To assist in improving the stability, FGF-1 and FGF-2 were connected via a 12-residue glycine linker ultimately producing a heterodimer. This heterodimer was further examined to discover its novel properties. Inspiration for this project was drawn from previous research which performed five mutations on FGF-1 ultimately improving stability in the protein complex. In addition to these five mutations on the FGF-1, sequence homology highlighted six residues on FGF-2 that could improve stability. Working with the second mutation on FGF-2 or the seventh mutation total, the heptamutant was a point mutation of K119N where a positively charged lysine was exchanged with a polar uncharged asparagine. The goal of this mutation was to reduce the overall positive charge on a region of the FGF-2 molecule called the heparin-binding pocket via a substitution. This mutation was believed to reduce the electrostatic strain, thus stabilizing the molecule. The structures and stability were tested to determine the impact of this mutation on the heterodimer. It was found that the mutation did not impact the secondary or tertiary structures. The Differential Scanning Calorimetry for the heptamutant demonstrated an increase in Tm of 7.61 °C compared to the wild-type. Similarly, a urea denaturation test showed an increase of Cm of approximately 1 M. Overall, it was found that the mutation increased stability without compromising a structural change showing promising signs for further endeavors towards wound healing applications.
Keywords
Fibroblast Growth Factor, FGF, Cell Proliferation, Angiogenesis, FGF-1-FGF-2 Heterodimer
Citation
Pohlman, N. A. (2023). Design of an FGF-1-FGF-2 Heterodimer Variant with Enhanced Stability and Cell Proliferation Activity. Chemistry & Biochemistry Undergraduate Honors Theses Retrieved from https://scholarworks.uark.edu/chbcuht/49
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