Reprogramming FGF1 from the natural growth factor to the engineered heparan sulphate biosensor
Abstract Heparan sulphate proteoglycans (HSPGs) are cell surface and extracellular matrix proteoglycan family members, playing a key role in diverse biological activities including signal transduction or endocytosis. To date, many HSPG ligands have been identified, including growth factors interacti...
Saved in:
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2025-05-01
|
| Series: | Cell Communication and Signaling |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12964-025-02269-x |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1850231205388091392 |
|---|---|
| author | Krzysztof Ciura Olimpia Sobota Aleksandra Chorążewska Daniel Krowarsch Natalia Porębska Łukasz Opaliński |
| author_facet | Krzysztof Ciura Olimpia Sobota Aleksandra Chorążewska Daniel Krowarsch Natalia Porębska Łukasz Opaliński |
| author_sort | Krzysztof Ciura |
| collection | DOAJ |
| description | Abstract Heparan sulphate proteoglycans (HSPGs) are cell surface and extracellular matrix proteoglycan family members, playing a key role in diverse biological activities including signal transduction or endocytosis. To date, many HSPG ligands have been identified, including growth factors interacting via sulfated domains of heparan sulphate (HS) chains. Due to significant overexpression of HSPGs in various cancers, discovering novel biomolecules capable of HSPGs recognition, which may act as HSPGs biosensors or drug carriers targeting HSPGs, is essential. Fibroblast growth factors (FGFs), mainly paracrine FGFs, are natural ligands of membrane-bound HSPGs, forming ternary signaling complex with HSPGs and fibroblast growth factor receptors (FGFRs). Here we employed the natural capability of FGF1 to bind HSPGs to develop HSPGs-specific proteinaceous probe. We identified three point mutations within FGF1 that elevate its affinity for heparin—S116R, S17K and L72R, named B, C and D respectively. Together with substitutions increasing FGF1 stability and abolishing FGF1 binding to FGFRs, we have generated eight HSPGs-specific variants. Among tested mutants, FGF1HSBCD exhibits the highest affinity for heparin and HS/HSPGs, showing over 20-fold increase in affinity for glypican-4 and requiring 0.23 M higher salt concentration for elution from heparin column compared to the initial FGF1HS molecule. Finally, we demonstrated FGF1HSBCD potential to act as a molecular sensor of HSPGs level in cancer cell lines overproducing HSPGs, implicating that FGF1HSBCD can be used as HSPGs biosensor or as a drug delivery carrier in protein-drug conjugates (PDC) targeting HSPGs. |
| format | Article |
| id | doaj-art-7c498fc407854fbaad468f76814202cb |
| institution | OA Journals |
| issn | 1478-811X |
| language | English |
| publishDate | 2025-05-01 |
| publisher | BMC |
| record_format | Article |
| series | Cell Communication and Signaling |
| spelling | doaj-art-7c498fc407854fbaad468f76814202cb2025-08-20T02:03:36ZengBMCCell Communication and Signaling1478-811X2025-05-0123111310.1186/s12964-025-02269-xReprogramming FGF1 from the natural growth factor to the engineered heparan sulphate biosensorKrzysztof Ciura0Olimpia Sobota1Aleksandra Chorążewska2Daniel Krowarsch3Natalia Porębska4Łukasz Opaliński5Department of Medical Biotechnology, Faculty of Biotechnology, University of WroclawDepartment of Medical Biotechnology, Faculty of Biotechnology, University of WroclawDepartment of Medical Biotechnology, Faculty of Biotechnology, University of WroclawDepartment of Protein Engineering, Faculty of Biotechnology, University of WroclawDepartment of Medical Biotechnology, Faculty of Biotechnology, University of WroclawDepartment of Medical Biotechnology, Faculty of Biotechnology, University of WroclawAbstract Heparan sulphate proteoglycans (HSPGs) are cell surface and extracellular matrix proteoglycan family members, playing a key role in diverse biological activities including signal transduction or endocytosis. To date, many HSPG ligands have been identified, including growth factors interacting via sulfated domains of heparan sulphate (HS) chains. Due to significant overexpression of HSPGs in various cancers, discovering novel biomolecules capable of HSPGs recognition, which may act as HSPGs biosensors or drug carriers targeting HSPGs, is essential. Fibroblast growth factors (FGFs), mainly paracrine FGFs, are natural ligands of membrane-bound HSPGs, forming ternary signaling complex with HSPGs and fibroblast growth factor receptors (FGFRs). Here we employed the natural capability of FGF1 to bind HSPGs to develop HSPGs-specific proteinaceous probe. We identified three point mutations within FGF1 that elevate its affinity for heparin—S116R, S17K and L72R, named B, C and D respectively. Together with substitutions increasing FGF1 stability and abolishing FGF1 binding to FGFRs, we have generated eight HSPGs-specific variants. Among tested mutants, FGF1HSBCD exhibits the highest affinity for heparin and HS/HSPGs, showing over 20-fold increase in affinity for glypican-4 and requiring 0.23 M higher salt concentration for elution from heparin column compared to the initial FGF1HS molecule. Finally, we demonstrated FGF1HSBCD potential to act as a molecular sensor of HSPGs level in cancer cell lines overproducing HSPGs, implicating that FGF1HSBCD can be used as HSPGs biosensor or as a drug delivery carrier in protein-drug conjugates (PDC) targeting HSPGs.https://doi.org/10.1186/s12964-025-02269-xFGF1FGFRHSPGHeparinHeparan sulfateProtein engineering |
| spellingShingle | Krzysztof Ciura Olimpia Sobota Aleksandra Chorążewska Daniel Krowarsch Natalia Porębska Łukasz Opaliński Reprogramming FGF1 from the natural growth factor to the engineered heparan sulphate biosensor Cell Communication and Signaling FGF1 FGFR HSPG Heparin Heparan sulfate Protein engineering |
| title | Reprogramming FGF1 from the natural growth factor to the engineered heparan sulphate biosensor |
| title_full | Reprogramming FGF1 from the natural growth factor to the engineered heparan sulphate biosensor |
| title_fullStr | Reprogramming FGF1 from the natural growth factor to the engineered heparan sulphate biosensor |
| title_full_unstemmed | Reprogramming FGF1 from the natural growth factor to the engineered heparan sulphate biosensor |
| title_short | Reprogramming FGF1 from the natural growth factor to the engineered heparan sulphate biosensor |
| title_sort | reprogramming fgf1 from the natural growth factor to the engineered heparan sulphate biosensor |
| topic | FGF1 FGFR HSPG Heparin Heparan sulfate Protein engineering |
| url | https://doi.org/10.1186/s12964-025-02269-x |
| work_keys_str_mv | AT krzysztofciura reprogrammingfgf1fromthenaturalgrowthfactortotheengineeredheparansulphatebiosensor AT olimpiasobota reprogrammingfgf1fromthenaturalgrowthfactortotheengineeredheparansulphatebiosensor AT aleksandrachorazewska reprogrammingfgf1fromthenaturalgrowthfactortotheengineeredheparansulphatebiosensor AT danielkrowarsch reprogrammingfgf1fromthenaturalgrowthfactortotheengineeredheparansulphatebiosensor AT nataliaporebska reprogrammingfgf1fromthenaturalgrowthfactortotheengineeredheparansulphatebiosensor AT łukaszopalinski reprogrammingfgf1fromthenaturalgrowthfactortotheengineeredheparansulphatebiosensor |