Lectins are carbohydrate-binding proteins ubiquitously present in nature. They play a role in biological recognition phenomena involving cells and proteins. The interaction lectin-carbohydrate is highly specific, and can be exploited for the development of nanoparticles containing on their surface lectins specifically directed to carbohydrate residues present only on malignant cells and absent on healthy ones [1]. Lectins have been found to possess anticancer properties and they are proposed as therapeutic agents, binding to cancer cell membranes or their receptors, causing cytotoxicity, apoptosis and inhibition of tumor growth. Some lectins are able to prevent the proliferation of malignant tumor cells because they recognize the T-antigen (Gal β 1–3GalNAc) found specifically on the surface of tumor cells [2]. The main problem is that their use as a detection agent for the T-antigen in clinical studies is not possible because the immune system can recognize them as foreign molecules and develop an immune response. Previous studies with X-ray crystallography made in our laboratory have characterized a lectin found in mushrooms called BEL β-trefoil which has antiproliferative activity on tumor cell lines, because it contains three binding sites for the T-antigen. Unlike other lectins with this property, BEL β-trefoil shows structural homology with a human protein, acidic Fibroblast Growth Factor (FGF1) [3]. Superposition of their structures suggests that the human protein could be mutated to contain at least one of the binding sites for the T-antigen. Such mutations should create in FGF1 the potential capacity of recognizing tumor cells with less immunogenicity than the fungal protein. FGF1 is mitogenic and chemotactic, and mediates cellular functions by binding to transmembrane receptors, which are activated by ligand-induced dimerization requiring heparin as co-receptor. To reach our purpose, the FGF1 cDNA was cloned into a bacterial plasmid and then mutated in four different positions to eliminate its mitogenic activity and to engineer in the protein the T-antigen binding capacity. Attempts to crystalize the mutants of FGF1 were made using the hanging drop technique with the final aim to carry out their structural characterization by X-ray diffraction analysis of the crystals. [1] Lis H and Sharon N. Lectins as molecules and as tools. Annu Rev Biochem. 1986. 55(1): p. 35-67. [2] Ju T, Otto VI, Cummings RD. The Tn antigen-structural simplicity and biological complexity. Angew Chem Int Ed Engl. 2011. 50(8): p.1770-1791. [3] Bovi M, Cenci L, Perduca M, Capaldi S, Carrizo ME, Civiero L, Chiarelli LR, Galliano M, Monaco HL. BEL β-trefoil: a novel lectin with antineoplastic properties in king bolete (Boletus edulis) mushrooms. 2013. Glycobiology. 23(5): p. 578-592. 89

Structural studies of human acidic fibroblast-growth factor (FGF1) mutants with a probable anticancer activity

GONZALEZ, Maria Cecilia;CAPALDI, Stefano;Destefanis, Laura;BOVI, Michele;PERDUCA, Massimiliano;MONACO, Ugo Luigi
2015-01-01

Abstract

Lectins are carbohydrate-binding proteins ubiquitously present in nature. They play a role in biological recognition phenomena involving cells and proteins. The interaction lectin-carbohydrate is highly specific, and can be exploited for the development of nanoparticles containing on their surface lectins specifically directed to carbohydrate residues present only on malignant cells and absent on healthy ones [1]. Lectins have been found to possess anticancer properties and they are proposed as therapeutic agents, binding to cancer cell membranes or their receptors, causing cytotoxicity, apoptosis and inhibition of tumor growth. Some lectins are able to prevent the proliferation of malignant tumor cells because they recognize the T-antigen (Gal β 1–3GalNAc) found specifically on the surface of tumor cells [2]. The main problem is that their use as a detection agent for the T-antigen in clinical studies is not possible because the immune system can recognize them as foreign molecules and develop an immune response. Previous studies with X-ray crystallography made in our laboratory have characterized a lectin found in mushrooms called BEL β-trefoil which has antiproliferative activity on tumor cell lines, because it contains three binding sites for the T-antigen. Unlike other lectins with this property, BEL β-trefoil shows structural homology with a human protein, acidic Fibroblast Growth Factor (FGF1) [3]. Superposition of their structures suggests that the human protein could be mutated to contain at least one of the binding sites for the T-antigen. Such mutations should create in FGF1 the potential capacity of recognizing tumor cells with less immunogenicity than the fungal protein. FGF1 is mitogenic and chemotactic, and mediates cellular functions by binding to transmembrane receptors, which are activated by ligand-induced dimerization requiring heparin as co-receptor. To reach our purpose, the FGF1 cDNA was cloned into a bacterial plasmid and then mutated in four different positions to eliminate its mitogenic activity and to engineer in the protein the T-antigen binding capacity. Attempts to crystalize the mutants of FGF1 were made using the hanging drop technique with the final aim to carry out their structural characterization by X-ray diffraction analysis of the crystals. [1] Lis H and Sharon N. Lectins as molecules and as tools. Annu Rev Biochem. 1986. 55(1): p. 35-67. [2] Ju T, Otto VI, Cummings RD. The Tn antigen-structural simplicity and biological complexity. Angew Chem Int Ed Engl. 2011. 50(8): p.1770-1791. [3] Bovi M, Cenci L, Perduca M, Capaldi S, Carrizo ME, Civiero L, Chiarelli LR, Galliano M, Monaco HL. BEL β-trefoil: a novel lectin with antineoplastic properties in king bolete (Boletus edulis) mushrooms. 2013. Glycobiology. 23(5): p. 578-592. 89
Human acidic fibroblast-growth factor, Anticancer activity, Lectins
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Descrizione: Structural studies of human acidic fibroblast-growth factor (FGF1) mutants with a probable anticancer activity
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11562/927069
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