Date of Graduation

5-2023

Document Type

Thesis

Degree Name

Bachelor of Science in Chemistry

Degree Level

Undergraduate

Department

Chemistry & Biochemistry

Advisor/Mentor

Thallapuranam, Suresh

Committee Member/Reader

Dong, Bin

Committee Member/Second Reader

Alrubaye, Adnan

Committee Member/Third Reader

Churchill, Hugh

Abstract

Fibroblast growth factors are a family of cell signaling proteins that have the ability to bind to heparin and are involved in a wide range of physiological functions, such as angiogenesis, cell differentiation, cell proliferation, cell morphogenesis, and wound healing. Fibroblast growth factor (FGF) 19 is a signaling protein belonging to this large family of fibroblast growth factors (FGFs). FGF19 has a pronounced effect on various biological processes, including cell proliferation, cholesterol and lipid catabolism, and glucose homeostasis. As such, FGF19 is known to play an integral role in regulating many metabolic diseases, which piques the interest of the science community. In this study, we have gained a better understanding of the characteristics regarding the secondary and tertiary structures of FGF19 as well as the thermal stability of FGF19 so as to achieve a better understanding of the mitogenic activities and metabolic activities as they have vast implications on glucose uptake activity. We did this by ascertaining the optimal parameters to over-express the FGF19 protein using E. coli and, subsequently, using nickel-Sepharose column chromatography for purification of the protein. That was then followed by a series of characterization studies to include secondary structure study of FGF19 using circular dichroism, which showed that FGF19 had no alterations in its secondary structure of primarily alpha helices, and the tertiary structure study using fluorescence spectroscopy indicated that the tertiary structure was also intact upon purification. The thermal stability of FGF19 was investigated using differential scanning calorimetry, which indicated a higher melting point for FGF19, which confers relatively low thermal stability. Future studies will further investigate the effects of FGF19 on the oxygen consumption rate of cells and on the glucose uptake activity in cells.

Keywords

Fibroblast Growth Factors; protein characterization; thermal stability

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