Three-dimensional structure of acidic fibroblast growth factor in solution: effects of binding to a heparin functional analog.
Acidic and basic fibroblast growth factors (aFGF and bFGF; FGFs) are paradigms of a group of nine closely related proteins known as the fibroblast growth factor family. FGFs induce mitosis in most mesoderm- and neuroectoderm-derived cells, and appear to be involved in diseases caused by anomalous cell proliferation. In vitro assays show that binding to heparin-like glycosaminoglycans is required to elicit the mitogenic activity of these proteins. It has been shown that myo-inositol hexasulfate (MIHS) emulates heparin in the mitogenesis assays of aFGF, and a low-resolution three-dimensional structure in solution of this protein bound to MIHS has been reported. Here we describe the 1H-NMR three-dimensional structure in solution of the free aFGF. Comparison of this structure with that of the protein bound to MIHS, upgraded to a level of refinement equivalent to that of the free protein, shows that MIHS binding causes some slight conformational changes with an increase in the definition of the structure. In addition, amide exchange H/2H rates of the most protected protons, and exchange data of the intermediate and fast-exchanging ones show that the free protein is less stable (< or = 2 kcal/mol) and more flexible in terms of local unfolding equilibria, respectively, than the MIHS-bound one. Thus, MIHS binding to aFGF causes a decrease of its flexibility, which translates into an enhancement of the definition of its three-dimensional structure. The increase of aFGF rigidity affects regions that include those involved in recognizing the cell membrane receptor. Thus, our data suggest that enhancement of structural definition may play a key role in the modulation of the affinity of aFGF by its receptor, and, consequently, of its specific mitogenic activity.
Pineda-Lucena
A
A
Centro de Investigaciones Biológicas (C.S.I.C.) Velázquez, Madrid, Spain.
Jiménez
M A
MA
Lozano
R M
RM
Nieto
J L
JL
Santoro
J
J
Rico
M
M
Giménez-Gallego
G
G
D000577:Amides / Q000737:chemistry; D016220:Fibroblast Growth Factor 1 / Q000737:chemistry* / Q000378:metabolism; D006493:Heparin / Q000031:analogs & derivatives / Q000378:metabolism; D006801:Humans; D006860:Hydrogen Bonding; D066298:In Vitro Techniques; D007294:Inositol / Q000031:analogs & derivatives / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D008958:Models, Molecular; D015394:Molecular Structure; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D011522:Protons; D012996:Solutions; D013816:Thermodynamics
D000577:Amides; D010446:Peptide Fragments; D011522:Protons; D012996:Solutions; D016220:Fibroblast Growth Factor 1; C013063:inositol hexasulfate; D007294:Inositol; D006493:Heparin