Computer‐assisted engineering of hyperstable fibroblast growth factor 2. Issue 4 (24th January 2018)

• Record Type: Journal Article
• Authors: Dvorak, Pavel; Bednar, David; Vanacek, Pavel; Balek, Lukas; Eiselleova, Livia; Stepankova, Veronika; Sebestova, Eva; Kunova Bosakova, Michaela; Konecna, Zaneta; Mazurenko, Stanislav; Kunka, Antonin; Vanova, Tereza; Zoufalova, Karolina; Chaloupkova, Radka; Brezovsky, Jan; Krejci, Pavel; Prokop, Zbynek; Dvorak, Petr; Damborsky, Jiri
• Is Part Of: Biotechnology and bioengineering. Volume 115:Issue 4(2018)
• Page Start: 850
• Page End: 862
• Publication Date: 2018-01-24
• Subjects: computational design -- embryonic stem cells -- fibroblast growth factor -- focused directed evolution -- protein engineering -- protein stability; Biotechnology -- Periodicals; Bioengineering -- Periodicals; 660.6
• DOI: 10.1002/bit.26531
• Languages: English
• ISSNs: 0006-3592

Abstract

Abstract: Fibroblast growth factors (FGFs) serve numerous regulatory functions in complex organisms, and their corresponding therapeutic potential is of growing interest to academics and industrial researchers alike. However, applications of these proteins are limited due to their low stability. Here we tackle this problem using a generalizable computer‐assisted protein engineering strategy to create a unique modified FGF2 with nine mutations displaying unprecedented stability and uncompromised biological function. The data from the characterization of stabilized FGF2 showed a remarkable prediction potential of in silico methods and provided insight into the unfolding mechanism of the protein. The molecule holds a considerable promise for stem cell research and medical or pharmaceutical applications. Abstract : The applications of fibroblast growth factors FGFs for stem cells cultivation and protein therapies are often limited by low stability of these proteins. Here the authors describe engineering hyperstable FGF2 molecule with melting temperature increased by 19 °C and in vitro functional half‐life at 37 °C improved from 10 hr to more than 20 days. This stabilization was achieved by a computer‐assisted engineering strategy combining energy‐based and evolution‐based analyses with focused directed evolution and unique functional screening of novel variants.