Effect of extension of the heparin binding pocket on the structure, stability, and cell proliferation activity of the human acidic fibroblast growth factor.


Abstract

Acidic human fibroblast growth factor (hFGF1) plays a key role in cell growth and proliferation. Activation of the cell surface FGF receptor is believed to involve the glycosaminoglycan, heparin. However, the exact role of heparin is a subject of considerable debate. In this context, in this study, the correlation between heparin binding affinity and cell proliferation activity of hFGF1 is examined by extending the heparin binding pocket through selective engineering via charge reversal mutations (D82R, D84R and D82R/D84R). Results of biophysical experiments such as intrinsic tryptophan fluorescence and far UV circular dichroism spectroscopy suggest that the gross native structure of hFGF1 is not significantly perturbed by the engineered mutations. However, results of limited trypsin digestion and ANS binding experiments show that the backbone structure of the D82R variant is more flexible than that of the wild type hFGF1. Results of the temperature and urea-induced equilibrium unfolding experiments suggest that the stability of the charge-reversal mutations increases in the presence of heparin. Isothermal titration calorimetry (ITC) data reveal that the heparin binding affinity is significantly increased when the charge on D82 is reversed but not when the negative charge is reversed at both positions D82 and D84 (D82R/D84R). However, despite the increased affinity of D82R for heparin, the cell proliferation activity of the D82R variant is observed to be reduced compared to the wild type hFGF1. The results of this study clearly demonstrate that heparin binding affinity of hFGF1 is not strongly correlated to its cell proliferation activity.

Submission Details

ID: jXbeHAoc3

Submitter: Shu-Ching Ou

Submission Date: March 28, 2019, 12:27 p.m.

Version: 1

Publication Details
Davis JE;Gundampati RK;Jayanthi S;Anderson J;Pickhardt A;Koppolu BP;Zaharoff DA;Kumar TKS,Biochem Biophys Rep (2018) Effect of extension of the heparin binding pocket on the structure, stability, and cell proliferation activity of the human acidic fibroblast growth factor. PMID:29556563
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
1AFC 1993-07-13T00:00:00+0000 2.7 STRUCTURAL STUDIES OF THE BINDING OF THE ANTI-ULCER DRUG SUCROSE OCTASULFATE TO ACIDIC FIBROBLAST GROWTH FACTOR
1BAR 1992-09-29T00:00:00+0000 2.7 THREE-DIMENSIONAL STRUCTURES OF ACIDIC AND BASIC FIBROBLAST GROWTH FACTORS
2J3P 2006-08-22T00:00:00+0000 1.4 crystal structure of rat FGF1 at 1.4 A
2UUS 2007-03-07T00:00:00+0000 2.2 Crystal structure of the rat FGF1-sucrose octasulfate (SOS) complex.
1AXM 1997-10-16T00:00:00+0000 3.0 HEPARIN-LINKED BIOLOGICALLY-ACTIVE DIMER OF FIBROBLAST GROWTH FACTOR
1DJS 1999-12-03T00:00:00+0000 2.4 LIGAND-BINDING PORTION OF FIBROBLAST GROWTH FACTOR RECEPTOR 2 IN COMPLEX WITH FGF1
1DZC 2000-02-24T00:00:00+0000 0 High resolution structure of acidic fibroblast growth factor. Mutant FGF-4-ALA-(24-154), 24 NMR structures
1DZD 2000-02-24T00:00:00+0000 0 High resolution structure of acidic fibroblast growth factor (27-154), 24 NMR structures
1E0O 2000-04-03T00:00:00+0000 2.8 CRYSTAL STRUCTURE OF A TERNARY FGF1-FGFR2-HEPARIN COMPLEX
1EVT 2000-04-20T00:00:00+0000 2.8 CRYSTAL STRUCTURE OF FGF1 IN COMPLEX WITH THE EXTRACELLULAR LIGAND BINDING DOMAIN OF FGF RECEPTOR 1 (FGFR1)

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
100.0 Fibroblast growth factor 1 P05230 FGF1_HUMAN
100.0 Fibroblast growth factor 1 Q5NVQ3 FGF1_PONAB
97.8 Fibroblast growth factor 1 P34004 FGF1_MESAU
96.4 Fibroblast growth factor 1 P61148 FGF1_MOUSE
96.4 Fibroblast growth factor 1 P61149 FGF1_RAT
97.8 Fibroblast growth factor 1 P20002 FGF1_PIG
92.1 Fibroblast growth factor 1 P03968 FGF1_BOVIN
90.6 Fibroblast growth factor 1 Q7M303 FGF1_SHEEP
90.6 Fibroblast growth factor 1 P19596 FGF1_CHICK
91.5 Fibroblast growth factor 1 Q9N1S8 FGF1_CAPCA