title: Effects of pH and polyanions on the thermal stability of fibroblast growth factor 20.
abstract: Fibroblast growth factor 20 (FGF20) is a member of the FGF family with potential for use in several different therapeutic categories. In this work, we provide the first structural characterization of FGF20 using a wide variety of approaches. Like other members of the FGF family, FGF20 appears to possess a beta-trefoil structure. The effect of pH on the conformation and thermal stability of FGF20 is evaluated using far-UV circular dichroism (CD), intrinsic and ANS fluorescence, and high-resolution derivative UV absorption spectroscopy. Empirical phase diagrams are constructed to describe the solution behavior of FGF20 over a wide pH and temperature range. The protein appears to be unstable at pH <5, with aggregation and precipitation observed during dialysis. A major heat-induced conformational change also causes aggregation and precipitation of FGF20 at elevated temperatures. The highest thermal stability is observed near neutral pH (Tm ~55 degrees C at pH 7). The effect of several high- and low-molecular mass polyanions on the thermal stability of FGF20 is also examined using CD, intrinsic fluorescence, and DSC analysis. Among these ligands, heparin exhibits the greatest stabilizing effect on FGF20, increasing the Tm by more than 10 degrees C.
authors.affiliation: Department of Pharmaceutical Chemistry, University of Kansas, Lawrence, KS 66044, USA.
authors.forename: Haihong
authors.initials: H
authors.lastname: Fan
authors.forename: Samadhi N
authors.initials: SN
authors.lastname: Vitharana
authors.forename: Tracy
authors.initials: T
authors.lastname: Chen
authors.forename: Donald
authors.initials: D
authors.lastname: O'Keefe
authors.forename: C Russell
authors.initials: CR
authors.lastname: Middaugh
mesh_terms: D002942:Circular Dichroism; D004355:Drug Stability; D005346:Fibroblast Growth Factors / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D000071228:Polyelectrolytes; D011108:Polymers / Q000494:pharmacology*; D011487:Protein Conformation; D011489:Protein Denaturation; D013050:Spectrometry, Fluorescence; D013696:Temperature
chemical_list: D005346:Fibroblast Growth Factors; D000071228:Polyelectrolytes; D011108:Polymers; C416003:FGF20 protein, human