Understanding the Structural Determinants for the Stability of Human Fibroblast Growth Factor

Human fibroblast growth factor (FGF) is an important protein that plays a role in morphogenesis, angiogenesis and wound healing. Because of these capabilities this protein is of medical interest. Unfortunately the lack of stability of FGF poses a problem to further research. Understanding the structure of FGF at the atomic level is necessary for designing novel FGF variants with enhanced stability and wound healing properties. The specific aim of this study is to test and record the structural stability of FGF through a gradient of salt concentrations. Through thermal denaturation, it was shown that the stability increased slightly with the increased salt concentration. The structure of hFGF-1 has been investigated using a variety of biophysical techniques such as trypsin digestion and circular dichroism. To ensure that the structure was maintained, N-15 enriched human fibroblast growth factor-1 was over expressed and purified. This sample will be used to test the structural stability of hFGF-1 by Heteronuclear Single Quantum Coherence (HSQC) using NMR spectroscopy. The hope of this study is to identify key properties that will allow the optimization of FGF stability. These data can then be used to develop novel mutants with enhanced stability and wound healing properties.