Highly stable mutants of human fibroblast growth factor-1 exhibit prolonged biological action.


Fibroblast growth factor 1 (FGF-1) shows strong angiogenic, osteogenic and tissue-injury repair properties that might be relevant to medical applications. Since FGF-1 is partially unfolded at physiological temperature we decided to increase significantly its conformational stability and test how such an improvement will affect its biological function. Using an homology approach and rational strategy we designed two new single FGF-1 mutations: Q40P and S47I that appeared to be the most strongly stabilizing substitutions among those reported so far, increasing the denaturation temperature by 7.8 deg. C and 9.0 deg. C, respectively. As our goal was to produce highly stable variants of the growth factor, we combined these two mutations with five previously described stabilizing substitutions. The multiple mutants showed denaturation temperatures up to 27 deg. C higher than the wild-type and exhibited full additivity of the mutational effects. All those mutants were biologically competent in several cell culture assays, maintaining typical FGF-1 activities, such as binding to specific cell surface receptors and activation of downstream signaling pathways. Thus, we demonstrate that the low denaturation temperature of wild-type FGF-1 is not related to its fundamental cellular functions, and that FGF-1 action is not affected by its stability. A more detailed analysis of the biological behavior of stable FGF-1 mutants revealed that, compared with the wild-type, their mitogenic properties, as probed by the DNA synthesis assay, were significantly increased in the absence of heparin, and that their half-lives were extensively prolonged. We found that the biological action of the mutants was dictated by their susceptibility to proteases, which strongly correlated with the stability. Mutants which were much more resistant to proteolytic degradation always displayed a significant improvement in the half-life and mitogenesis. Our results show that engineered stable growth factor variants exhibit enhanced and prolonged activity, which can be advantageous in terms of the potential therapeutic applications of FGF-1.

Submission Details

ID: 4j4tndHa

Submitter: Shu-Ching Ou

Submission Date: Sept. 29, 2018, 10:06 a.m.

Version: 1

Publication Details
Zakrzewska M;Krowarsch D;Wiedlocha A;Olsnes S;Otlewski J,J Mol Biol (2005) Highly stable mutants of human fibroblast growth factor-1 exhibit prolonged biological action. PMID:16126225
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 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
97.9 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
90.7 Fibroblast growth factor 1 Q7M303 FGF1_SHEEP
91.5 Fibroblast growth factor 1 Q9N1S8 FGF1_CAPCA