Date of Graduation

12-2023

Document Type

Thesis

Degree Name

Master of Science in Cell & Molecular Biology (MS)

Degree Level

Graduate

Department

Biological Sciences

Advisor/Mentor

Suresh Thallapuranam Kumar

Committee Member

Sakon, Joshua

Second Committee Member

Ceballos, Ruben Michael

Keywords

FGF2, Fibroblast growth factors

Abstract

Fibroblast growth factors (FGFs) are a family of cell signaling proteins conserved across multiple species. Each individual FGF elicits different cellular functions including, but not limited to, proliferation, migration, differentiation, angiogenesis, and wound healing. One of the most studied members, fibroblast growth factor 2 (FGF2), has demonstrated substantial wound healing capacity in a wide range of tissues including skeletal, muscular, neural, respiratory, epithelial, and cardiovascular. This ability makes FGF2 a potential therapeutic for a wide range of conditions and injuries. However, due to a short half-life at room temperature, therapeutic use of FGF2 is limited. It has been demonstrated that in the structurally similar FGF1, point mutations in the heparin binding region significantly improved the proteins’ stability and mitogenic activity. These mutations decreased the overall positive charge in the heparin binding region. Similar mutations in FGF2 may produce similar effects, since there are several positive residues in the heparin binding region of FGF2. Other research has identified lysine 134 as an important residue of the heparin binding region and was thereby chosen for charge reverse mutation. This research designed a K134E mutant of FGF2, and investigated the effects on stability and mitogenic activity. Using various biophysical techniques, proteolytic assay, and cell proliferation assay, we demonstrated that the engineered K134E mutant displayed enhanced bioactivity and thermal stability.

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