A combination of near- and far-UV circular dichroism, Fourier-transform infrared spectroscopy, tryptophan fluorescence, size-exclusion chromatography, and a fluorescent extrinsic hydrophobic probe has been employed to characterize partially structured states of human recombinant acidic fibroblast growth factor (aFGF). At low pH, the addition of specific polyanionic ligands or moderate amounts of salts induces states with high secondary but low tertiary structure content. At neutral pH, intermediate amounts of chaotropic agents impose similar partially structured conformational states which also display noncooperative unfolding transitions. Kinetic evidence indicates that similar forms of the protein exist in the first few hundred milliseconds in the refolding pathway of aFGF. The kinetics of their formation appear to be temperature-independent, implying lack of an energy barrier, which is characteristic for further slow folding into the native state. Unlike the native and fully unfolded states, these partially structured conformations exhibit very low solubility, resulting in irreversible aggregation. Potential physiological implications of the existence of such "molten globule" states with regard to the growth factor's transport and biological activity are considered. Study holds ProTherm entries: 4561 Extra Details: additive : EDTA(0.1 mM), tertiary structure; conformational states; noncooperative;,solubility; molten globule
ID: Kshaji3P
Submitter: Connie Wang
Submission Date: April 24, 2018, 8:26 p.m.
Version: 1
Number of data points | 2 |
Proteins | Fibroblast growth factor 2 ; Fibroblast growth factor 2 |
Unique complexes | 1 |
Assays/Quantities/Protocols | Experimental Assay: Cm ; Experimental Assay: dG_H2O |
Libraries | Mutations for sequence PALPEDGGSGAFPPGHFKDPKRLYCKNGGFFLRIHPDGRVDGVREKSDPHIKLQLQAEERGVVSIKGVCANRYLAMKEDGRLLASKCVTDECFFFERLESNNYNTYRSRKYTSWYVALKRTGQYKLGSKTGPGQKAILFLPMSAKS |
Colors: | D | E | R | H | K | S | T | N | Q | A | V | I | L | M | F | Y | W | C | G | P |
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Structure ID | Release Date | Resolution | Structure Title |
---|---|---|---|
1BAS | 1992-09-29T00:00:00+0000 | 1.9 | THREE-DIMENSIONAL STRUCTURES OF ACIDIC AND BASIC FIBROBLAST GROWTH FACTORS |
1BFB | 1995-12-12T00:00:00+0000 | 1.9 | BASIC FIBROBLAST GROWTH FACTOR COMPLEXED WITH HEPARIN TETRAMER FRAGMENT |
1BFC | 1995-12-12T00:00:00+0000 | 2.2 | BASIC FIBROBLAST GROWTH FACTOR COMPLEXED WITH HEPARIN HEXAMER FRAGMENT |
1BFF | 1996-12-06T00:00:00+0000 | 2.0 | THE 154 AMINO ACID FORM OF HUMAN BASIC FIBROBLAST GROWTH FACTOR |
1BFG | 1993-04-15T00:00:00+0000 | 1.6 | CRYSTAL STRUCTURE OF BASIC FIBROBLAST GROWTH FACTOR AT 1.6 ANGSTROMS RESOLUTION |
1BLA | 1996-05-20T00:00:00+0000 | 0 | BASIC FIBROBLAST GROWTH FACTOR (FGF-2) MUTANT WITH CYS 78 REPLACED BY SER AND CYS 96 REPLACED BY SER, NMR |
1BLD | 1996-05-20T00:00:00+0000 | 0 | BASIC FIBROBLAST GROWTH FACTOR (FGF-2) MUTANT WITH CYS 78 REPLACED BY SER AND CYS 96 REPLACED BY SER, NMR |
1CVS | 1999-08-24T00:00:00+0000 | 2.8 | CRYSTAL STRUCTURE OF A DIMERIC FGF2-FGFR1 COMPLEX |
1EV2 | 2000-04-19T00:00:00+0000 | 2.2 | CRYSTAL STRUCTURE OF FGF2 IN COMPLEX WITH THE EXTRACELLULAR LIGAND BINDING DOMAIN OF FGF RECEPTOR 2 (FGFR2) |
1FGA | 1993-02-26T00:00:00+0000 | 2.2 | REFINEMENT OF THE STRUCTURE OF HUMAN BASIC FIBROBLAST GROWTH FACTOR AT 1.6 ANGSTROMS RESOLUTION AND ANALYSIS OF PRESUMED HEPARIN BINDING SITES BY SELENATE SUBSTITUTION |
Percent Identity | Matching Chains | Protein | Accession | Entry Name |
---|---|---|---|---|
100.0 | Fibroblast growth factor 2 | P09038 | FGF2_HUMAN | |
100.0 | Fibroblast growth factor 2 | Q5IS69 | FGF2_PANTR | |
98.7 | Fibroblast growth factor 2 | P03969 | FGF2_BOVIN | |
98.1 | Fibroblast growth factor 2 | P20003 | FGF2_SHEEP | |
97.3 | Fibroblast growth factor 2 | P13109 | FGF2_RAT | |
96.6 | Fibroblast growth factor 2 | P15655 | FGF2_MOUSE | |
92.9 | Fibroblast growth factor 2 | P48798 | FGF2_MONDO | |
92.2 | Fibroblast growth factor 2 | P48800 | FGF2_CHICK | |
99.3 | Fibroblast growth factor 2 | P48799 | FGF2_RABIT | |
91.1 | Fibroblast growth factor 2 | Q60487 | FGF2_CAVPO |