title: Conformational flexibility, hydration and state parameter fluctuations of fibroblast growth factor-10: effects of ligand binding.
abstract: Differential effects of ligand binding on local and global fibroblast growth factor-10 (FGF-10) flexibility and stability have been investigated utilizing a variety of experimental and computational techniques. Normal mode analysis was used to predict the low frequency motions and regional flexibility of FGF-10. Similarly, regional variations in local folding/unfolding equilibria were characterized with the COREX/BEST algorithm. Experimental adiabatic and isothermal compressibilities of FGF-10 alone and in the presence of polyanions are compared. Furthermore, the effect of polyanions on the coefficient of thermal expansion is compared. Measurements of density, heat capacity, compressibility, and expansibility were combined to calculate experimentally determined volume and enthalpy fluctuations. Global effects of polyanions on FGF-10 flexibility, thermodynamic fluctuations, and hydration vary depending on the size and charge density of the polyanion. Local effects of polyanions were investigated utilizing time-resolved fluorescence spectroscopy and red edge excitation spectroscopy (REES). Increased rigidity of the protein matrix or an increased solvent response surrounding the Trp residues is observed in the presence of polyanions. Similarly, time-resolved spectroscopy reveals increased ground state heterogeneity and increased dipole relaxation on the time scale of fluorescence for FGF-10 in the presence of polyanions. These polyanions increase heterogeneity, global flexibility, and fluctuations while increasing the melting temperature (Tm) of FGF-10.
authors.affiliation: Department of Pharmaceutical Chemistry, University of Kansas, Lawrence, Kansas 66047, USA.
authors.forename: Tim J
authors.initials: TJ
authors.lastname: Kamerzell
authors.forename: Jay R
authors.initials: JR
authors.lastname: Unruh
authors.forename: Carey K
authors.initials: CK
authors.lastname: Johnson
authors.forename: C Russell
authors.initials: CR
authors.lastname: Middaugh
mesh_terms: D018360:Crystallography, X-Ray; D051526:Fibroblast Growth Factor 10 / Q000737:chemistry* / Q000378:metabolism*; D006801:Humans; D008024:Ligands; D000071228:Polyelectrolytes; D011108:Polymers / Q000378:metabolism; D011485:Protein Binding; D011487:Protein Conformation; D013050:Spectrometry, Fluorescence; D013053:Spectrophotometry; D013057:Spectrum Analysis; D013816:Thermodynamics; D014364:Tryptophan / Q000737:chemistry; D014867:Water / Q000737:chemistry*
chemical_list: D051526:Fibroblast Growth Factor 10; D008024:Ligands; D000071228:Polyelectrolytes; D011108:Polymers; D014364:Tryptophan; D014867:Water; C496463:FGF10 protein, human