Design of fully active FGF-1 variants with increased stability.


Abstract

Fibroblast growth factor 1 is a powerful mitogen playing an important role in morphogenesis, angiogenesis and wound healing and is therefore of potential medical interest. Using homologous sequence and structure comparisons, we designed and constructed 16 mutants of FGF-1 with increased thermodynamic stability, as determined by chemical and heat denaturation. For multiple mutants, additive effects on stability were observed, providing mutants up to 7.8 degrees C more stable than the wild-type. None of the introduced mutations affected any FGF-1 biological activities, such as stimulation of DNA synthesis, MAP kinase activation and binding to the FGF receptor on the cell surface. Our study provides a good starting point to improve the stability of FGF-1 in the context of its wide potential therapeutic applications. We showed that a homology approach is an effective method to change the thermodynamic properties of the protein without altering its function. Study holds ProTherm entries: 18239, 18240, 18241, 18242, 18243, 18244, 18245, 18246, 18247, 18248, 18249, 18250, 18251, 18252, 18253, 18254, 18255, 18256, 18257, 18258, 18259, 18260, 18261, 18262, 18263, 18264, 18265, 18266, 18267, 18268, 18269, 18270, 18271, 18272, 18273, 18274, 18275, 18276, 18277, 18278, 18279, 18280, 18281, 18282, 18283, 18284, 18285, 18286, 18287, 18288, 18289, 18290, 18291, 18292, 18293, 18294, 18295, 18296, 18297, 18298, 18299, 18300, 18301, 18302, 18303, 18304, 18305, 18306, 18307, 18308, 18309, 18310 Extra Details: DNA synthesis; fibroblast growth factor; MAP kinase activation; protein engineering; stability

Submission Details

ID: WBrHrZUr3

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:50 p.m.

Version: 1

Publication Details
Zakrzewska M;Krowarsch D;Wiedlocha A;Otlewski J,Protein Eng. Des. Sel. (2004) Design of fully active FGF-1 variants with increased stability. PMID:15469994
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
91.5 Fibroblast growth factor 1 Q9N1S8 FGF1_CAPCA
90.3 Fibroblast growth factor 1 P19596 FGF1_CHICK
90.7 Fibroblast growth factor 1 Q7M303 FGF1_SHEEP
92.3 Fibroblast growth factor 1 P03968 FGF1_BOVIN
96.4 Fibroblast growth factor 1 P61149 FGF1_RAT
96.4 Fibroblast growth factor 1 P61148 FGF1_MOUSE
98.0 Fibroblast growth factor 1 P20002 FGF1_PIG
97.9 Fibroblast growth factor 1 P34004 FGF1_MESAU
100.0 Fibroblast growth factor 1 Q5NVQ3 FGF1_PONAB
100.0 Fibroblast growth factor 1 P05230 FGF1_HUMAN