Physical stabilization of acidic fibroblast growth factor by polyanions.


Abstract

Acidic fibroblast growth factor (aFGF) is markedly stabilized by heparin. Partially due to the heterogeneity of heparin preparations, the nature of the aFGF polyanion binding site is still ill-defined. We have, therefore, investigated a wide variety of well-defined polyanions in terms of their ability to stabilize human recombinant aFGF (15-154) against thermal denaturation. The specificity of the interaction between aFGF and polyanions is shown to be remarkably weak with a surprising number of polyanions (including small phosphorylated and sulfated compounds as well as highly charged biopolymers) able to induce physical stability. Temperature-dependent fluorescence and circular dichroism measurements show that many of these polyanionic compounds stabilize aFGF to the same extent as heparin. The ability of these agents to protect the three free thiol groups of aFGF from copper-catalyzed oxidation was also explored and significant protection was observed. The extent and electrostatic requirements of the protein's polyanion binding site were probed by the use of a series of well-defined heparin fragments and differentially phosphorylated inositol compounds. A tetrasaccharide fragment of heparin is the smallest unit of heparin capable of stabilizing aFGF against thermal denaturation. Increasing phosphorylation of inositol compounds (up to six phosphate groups per molecule) enhances the thermal stability of aFGF. These results are discussed in the context of a model of human aFGF based on the X-ray crystal structure of the bovine protein and previous studies by others of the heparin binding site of both acidic and basic FGF. Study holds ProTherm entries: 7283 Extra Details:

Submission Details

ID: Co4trqay

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:33 p.m.

Version: 1

Publication Details
Volkin DB;Tsai PK;Dabora JM;Gress JO;Burke CJ;Linhardt RJ;Middaugh CR,Arch. Biochem. Biophys. (1993) Physical stabilization of acidic fibroblast growth factor by polyanions. PMID:7678726
Additional Information

Structure view and single mutant data analysis

Study data

No weblogo for data of varying length.
Colors: D E R H K S T N Q A V I L M F Y W C G P
 

Data Distribution

Studies with similar sequences (approximate matches)

Correlation with other assays (exact sequence matches)


Relevant PDB Entries

Structure ID Release Date Resolution Structure Title
1BLD 1996-11-08 BASIC FIBROBLAST GROWTH FACTOR (FGF-2) MUTANT WITH CYS 78 REPLACED BY SER AND CYS 96 REPLACED BY SER, NMR
2M49 2013-12-18 Structural Insights into Human S100B and Basic Fibroblast Growth Factor (FGF2) Interaction
1BLA 1996-11-08 BASIC FIBROBLAST GROWTH FACTOR (FGF-2) MUTANT WITH CYS 78 REPLACED BY SER AND CYS 96 REPLACED BY SER, NMR
4OEE 2014-07-09 1.5 Crystal Structure Analysis of FGF2-Disaccharide (S3I2) complex
4FGF 1993-07-15 1.6 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
1BFG 1994-01-31 1.6 CRYSTAL STRUCTURE OF BASIC FIBROBLAST GROWTH FACTOR AT 1.6 ANGSTROMS RESOLUTION
4OEG 2014-07-09 1.6 Crystal Structure Analysis of FGF2-Disaccharide (S9I2) complex
2FGF 1992-01-15 1.77 THREE-DIMENSIONAL STRUCTURE OF HUMAN BASIC FIBROBLAST GROWTH FACTOR, A STRUCTURAL HOMOLOG OF INTERLEUKIN 1BETA
4OEF 2014-07-09 1.8 Crystal Structure Analysis of FGF2-Disaccharide (S6I2) complex
5X1O 2018-03-07 1.9 PI(4,5)P2 lipid binding induced a reorientation of FGF2 molecules near membrane surface to facilitate the unconventional oligomerization-dependent secretion process as revealed by a combined FTIR/NMR/X-ray study
1BFB 1996-04-03 1.9 BASIC FIBROBLAST GROWTH FACTOR COMPLEXED WITH HEPARIN TETRAMER FRAGMENT
1BAS 1993-10-31 1.9 THREE-DIMENSIONAL STRUCTURES OF ACIDIC AND BASIC FIBROBLAST GROWTH FACTORS
1BFF 1997-06-16 2.0 THE 154 AMINO ACID FORM OF HUMAN BASIC FIBROBLAST GROWTH FACTOR
1EV2 2000-05-31 2.2 CRYSTAL STRUCTURE OF FGF2 IN COMPLEX WITH THE EXTRACELLULAR LIGAND BINDING DOMAIN OF FGF RECEPTOR 2 (FGFR2)
1BFC 1996-04-03 2.2 BASIC FIBROBLAST GROWTH FACTOR COMPLEXED WITH HEPARIN HEXAMER FRAGMENT
1FGA 1993-07-15 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
1IIL 2001-05-09 2.3 CRYSTAL STRUCTURE OF PRO253ARG APERT MUTANT FGF RECEPTOR 2 (FGFR2) IN COMPLEX WITH FGF2
2BFH 1994-01-31 2.5 CRYSTAL STRUCTURE OF BASIC FIBROBLAST GROWTH FACTOR AT 1.6 ANGSTROMS RESOLUTION
1II4 2001-05-09 2.7 CRYSTAL STRUCTURE OF SER252TRP APERT MUTANT FGF RECEPTOR 2 (FGFR2) IN COMPLEX WITH FGF2
1CVS 2000-01-28 2.8 CRYSTAL STRUCTURE OF A DIMERIC FGF2-FGFR1 COMPLEX
1FQ9 2000-09-27 3.0 CRYSTAL STRUCTURE OF A TERNARY FGF2-FGFR1-HEPARIN COMPLEX

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
99.3 Fibroblast growth factor 2 P48799 FGF2_RABIT
92.2 Fibroblast growth factor 2 P48800 FGF2_CHICK
92.9 Fibroblast growth factor 2 P48798 FGF2_MONDO
96.6 Fibroblast growth factor 2 P15655 FGF2_MOUSE
97.3 Fibroblast growth factor 2 P13109 FGF2_RAT
98.1 Fibroblast growth factor 2 P20003 FGF2_SHEEP
98.7 Fibroblast growth factor 2 P03969 FGF2_BOVIN
100.0 Fibroblast growth factor 2 Q5IS69 FGF2_PANTR
100.0 Fibroblast growth factor 2 P09038 FGF2_HUMAN
91.1 Fibroblast growth factor 2 Q60487 FGF2_CAVPO