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Stable Fibroblast Growth Factor 2 Dimers with High Pro-Survival and Mitogenic Potential

Fibroblast growth factor 2 (FGF2) is a heparin-binding growth factor with broad mitogenic and cell survival activities. Its effector functions are induced upon the formation of 2:2 FGF2:FGFR1 tetrameric complex. To facilitate receptor activation, and therefore, to improve the FGF2 biological propert...

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Autores principales: Nawrocka, Daria, Krzyscik, Mateusz Adam, Opaliński, Łukasz, Zakrzewska, Malgorzata, Otlewski, Jacek
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7312490/
https://www.ncbi.nlm.nih.gov/pubmed/32526859
http://dx.doi.org/10.3390/ijms21114108
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author Nawrocka, Daria
Krzyscik, Mateusz Adam
Opaliński, Łukasz
Zakrzewska, Malgorzata
Otlewski, Jacek
author_facet Nawrocka, Daria
Krzyscik, Mateusz Adam
Opaliński, Łukasz
Zakrzewska, Malgorzata
Otlewski, Jacek
author_sort Nawrocka, Daria
collection PubMed
description Fibroblast growth factor 2 (FGF2) is a heparin-binding growth factor with broad mitogenic and cell survival activities. Its effector functions are induced upon the formation of 2:2 FGF2:FGFR1 tetrameric complex. To facilitate receptor activation, and therefore, to improve the FGF2 biological properties, we preorganized dimeric ligand by a covalent linkage of two FGF2 molecules. Mutations of the FGF2 WT protein were designed to obtain variants with a single surface-exposed reactive cysteine for the chemical conjugation via maleimide-thiol reaction with bis-functionalized linear PEG linkers. We developed eight FGF2 dimers of defined topology, differing in mutual orientation of individual FGF2 molecules. The engineered proteins remained functional in terms of FGFR downstream signaling activation and were characterized by the increased stability, mitogenic potential and anti-apoptotic activity, as well as induced greater migration responses in normal fibroblasts, as compared to FGF2 monomer. Importantly, biological activity of the dimers was much less dependent on the external heparin administration. Moreover, some dimeric FGF2 variants internalized more efficiently into FGFR overexpressing cancer cells. In summary, in the current work, we showed that preorganization of dimeric FGF2 ligand increased the stability of the growth factor, and therefore, enhanced its biological activity.
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spelling pubmed-73124902020-06-29 Stable Fibroblast Growth Factor 2 Dimers with High Pro-Survival and Mitogenic Potential Nawrocka, Daria Krzyscik, Mateusz Adam Opaliński, Łukasz Zakrzewska, Malgorzata Otlewski, Jacek Int J Mol Sci Article Fibroblast growth factor 2 (FGF2) is a heparin-binding growth factor with broad mitogenic and cell survival activities. Its effector functions are induced upon the formation of 2:2 FGF2:FGFR1 tetrameric complex. To facilitate receptor activation, and therefore, to improve the FGF2 biological properties, we preorganized dimeric ligand by a covalent linkage of two FGF2 molecules. Mutations of the FGF2 WT protein were designed to obtain variants with a single surface-exposed reactive cysteine for the chemical conjugation via maleimide-thiol reaction with bis-functionalized linear PEG linkers. We developed eight FGF2 dimers of defined topology, differing in mutual orientation of individual FGF2 molecules. The engineered proteins remained functional in terms of FGFR downstream signaling activation and were characterized by the increased stability, mitogenic potential and anti-apoptotic activity, as well as induced greater migration responses in normal fibroblasts, as compared to FGF2 monomer. Importantly, biological activity of the dimers was much less dependent on the external heparin administration. Moreover, some dimeric FGF2 variants internalized more efficiently into FGFR overexpressing cancer cells. In summary, in the current work, we showed that preorganization of dimeric FGF2 ligand increased the stability of the growth factor, and therefore, enhanced its biological activity. MDPI 2020-06-09 /pmc/articles/PMC7312490/ /pubmed/32526859 http://dx.doi.org/10.3390/ijms21114108 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Nawrocka, Daria
Krzyscik, Mateusz Adam
Opaliński, Łukasz
Zakrzewska, Malgorzata
Otlewski, Jacek
Stable Fibroblast Growth Factor 2 Dimers with High Pro-Survival and Mitogenic Potential
title Stable Fibroblast Growth Factor 2 Dimers with High Pro-Survival and Mitogenic Potential
title_full Stable Fibroblast Growth Factor 2 Dimers with High Pro-Survival and Mitogenic Potential
title_fullStr Stable Fibroblast Growth Factor 2 Dimers with High Pro-Survival and Mitogenic Potential
title_full_unstemmed Stable Fibroblast Growth Factor 2 Dimers with High Pro-Survival and Mitogenic Potential
title_short Stable Fibroblast Growth Factor 2 Dimers with High Pro-Survival and Mitogenic Potential
title_sort stable fibroblast growth factor 2 dimers with high pro-survival and mitogenic potential
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7312490/
https://www.ncbi.nlm.nih.gov/pubmed/32526859
http://dx.doi.org/10.3390/ijms21114108
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