Computer-aided engineering of stabilized fibroblast growth factor 21
Gabin de La Bourdonnaye1,2, Tereza Ghazalova2, Petr Fojtik3
1Department of Experimental Biology, Faculty of Science, Masaryk University, Brno, Czech Republic.
Computational and Structural Biotechnology Journal
|February 21, 2024
Summary
Fibroblast growth factor 21 (FGF21) variants were engineered for enhanced stability and therapeutic potential in metabolic diseases. Computer-aided design improved thermostability without compromising efficacy in cell-based assays.
Area of Science:
- Biochemistry
- Molecular Biology
- Metabolic Engineering
Background:
- Fibroblast growth factor 21 (FGF21) is a key metabolic regulator with therapeutic potential for metabolic diseases.
- FGF21's therapeutic utility is limited by its poor stability (heat, acid, proteolysis) and rapid kidney clearance.
- Protein engineering strategies, particularly enhancing thermostability, have shown promise for improving FGF21 pharmacokinetics.
Purpose of the Study:
- To design and characterize FGF21 variants with improved thermostability using computer-aided design.
- To evaluate the efficacy of engineered FGF21 variants in cellular assays.
- To assess the potential of rational design in enhancing FGF21's pharmaceutical viability.
Main Methods:
- Computer-aided design (CAD) was employed to engineer FGF21 variants.
- Experimental characterization included measuring melting temperature (Tm) to assess thermostability.
- Cell-based assays (Hep G2, NIH 3T3 fibroblasts) were used to evaluate MAPK/ERK signaling activation and cell proliferation.
Main Results:
- Engineered FGF21 variants exhibited enhanced melting temperatures, up to 15°C higher than wild-type (WT).
- The FGF21 variants maintained uncompromised efficacy in activating MAPK/ERK signaling in Hep G2 cells.
- Stimulation of Hep G2 and NIH 3T3 fibroblast proliferation by the engineered variants was comparable to FGF21-WT.
Conclusions:
- Rational, computer-aided design can significantly enhance the thermostability of FGF21.
- Stabilized FGF21 variants retain biological efficacy, suggesting improved therapeutic potential.
- Combining rational design with other stabilization methods could maximize FGF21's pharmaceutical applications.
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