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Characterization of the Structure, Stability, Bioactivity of Recombinant FGF19
Author(s) -
Ternier Gaëtane,
Kumar T.K.S
Publication year - 2022
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2022.36.s1.r3100
Subject(s) - fgf19 , circular dichroism , fibroblast growth factor , fibroblast growth factor receptor 4 , chemistry , denaturation (fissile materials) , recombinant dna , biochemistry , protein folding , cell growth , fibroblast growth factor receptor , biophysics , biology , receptor , gene , nuclear chemistry
Fibroblast growth factor 19 (FGF19) is a member of the endocrine fibroblast growth factor family, secreted in the small intestine. FGF19 binds to the fibroblast growth factor receptor 4 (FGFR4) in the presence of β klotho to downregulate the secretion of bile acid. All endocrine FGFs, including FGF19 are unstable. In this study, we examine structure, stability, and bioactivity using a wide range of biophysical and biochemical experiments. The protein has been successfully overexpressed in Escherichia coli (E. coli), and optimal conditions for storage have been established. Results of thermal denaturation of recombinant FGF19, using circular‐dichroism and differential scanning calorimetry, suggest that the thermal unfolding of the protein is reversible, and the Tm (melting temperature; temperature at which half of the protein is denatured) was found to be 55 o C. Far UV circular‐dichroism data indicate that the backbone of the protein is predominantly in the beta‐sheet conformation. In addition, cell proliferation and cell metabolism data show that FGF19 exhibits low cell proliferation, but significant metabolic activity. These results will be discussed in details.