1H-NMR assignment and solution structure of human acidic fibroblast growth factor activated by inositol hexasulfate.
A major fragment of human acidic fibroblast growth factor of 132 amino acid residues is shown to be as active and stable as the 139 residue molecule initially described, and commonly used in physiological studies. It is shown that inositol hexasulfate is a good substitute for heparin in both activating and protecting acidic fibroblast growth factor. The complex between the shortened form of the protein and inositol hexasulfate was used to determine the structure of activated acidic fibroblast growth factor in solution. The 1H-NMR spectrum of the complex was totally assigned, and a low-resolution, three-dimensional structure of the protein computed. The global fold of the activated acidic fibroblast growth factor is similar to that proposed for a crystallized variant of the protein obtained by genetic engineering whose activity is not dependent on heparin. The inositol hexasulfate binds to the protein through the positively charged groups of Lys126, Lys127, Arg133 and Lys142 side-chains. The computed three-dimensional structure suggests that inositol hexasulfate may stabilize and activate the protein by conferring rigidity to the hairpin involving beta-strands 10 and 11.
Pineda-Lucena
A
A
Centro de Investigaciones Biológicas (C.S.I.C.), Madrid, Spain.
Jiménez
M A
MA
Nieto
J L
JL
Santoro
J
J
Rico
M
M
Giménez-Gallego
G
G
D000595:Amino Acid Sequence; D016220:Fibroblast Growth Factor 1 / Q000737:chemistry*; D006801:Humans; D006860:Hydrogen Bonding; D007294:Inositol / Q000031:analogs & derivatives* / Q000737:chemistry; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins
D011994:Recombinant Proteins; D016220:Fibroblast Growth Factor 1; C013063:inositol hexasulfate; D007294:Inositol