Poly-N-Acetyllactosamine Neo-Glycoproteins as Nanomolar Ligands of Human Galectin-3: Binding Kinetics and Modeling

Abstract

Galectin-3 (Gal-3) is recognized as a prognostic marker in several cancer types. Its involvement in tumor development and proliferation makes this lectin a promising target for early cancer diagnosis and anti-cancer therapies. Gal-3 recognizes poly-N-acetyllactosamine (LacNAc)-based carbohydrate motifs of glycoproteins and glycolipids with a high specificity for internal LacNAc epitopes. This study analyzes the mode and kinetics of binding of Gal-3 to a series of multivalent neo-glycoproteins presenting complex poly-LacNAc-based oligosaccharide ligands on a scaffold of bovine serum albumin. These neo-glycoproteins rank among the strongest Gal-3 ligands reported, with Kd reaching sub-nanomolar values as determined by surface plasmon resonance. Significant differences in the binding kinetics were observed within the ligand series, showing the tetrasaccharide capped with N,N′-diacetyllactosamine (LacdiNAc) as the strongest ligand of Gal-3 in this study. A molecular model of the Gal-3 carbohydrate recognition domain with docked oligosaccharide ligands is presented that shows the relations in the binding site at the molecular level. The neo-glycoproteins presented herein may be applied for selective recognition of Gal-3 both on the cell surface and in blood serum.

Sequence‐Defined Heteromultivalent Precision Glycomacromolecules Bearing Sulfonated/Sulfated Nonglycosidic Moieties Preferentially Bind Galectin‐3 and Delay Wound Healing of a Galectin‐3 Positive Tumor Cell Line in an In Vitro Wound Scratch Assay

“…ELISA‐type inhibition studies were performed according to an already established protocol by Elling and co‐workers. [ 30 ] Inhibition of Gal‐1 and Gal‐3 binding to asialofetuin was investigated for glycoligands 1b – 13b (and 8a for Gal‐1) with final concentrations between 0.1 × 10 −6 and 2000 × 10 −6 m using phosphate buffered saline (PBS) buffer (150 × 10 −3 m NaCl, 50 × 10 −3 m NaH 2 PO 4 , pH 7.5). All measurements were performed two times in triplicates.…”

“…[ 2,8–17 ] In cases where galectin‐3 has been shown to be responsible for tumor metastasis and migration, several groups have endeavored to design compounds to inhibit them. One group of examples involves the multivalent presentation of carbohydrate residues on macromolecular scaffolds as described by Gabius et al., [ 18,19 ] Roy et al., [ 20,21 ] Nilsson et al., [ 22,23 ] Wang et al., [ 24 ] Argueso et al., [ 25 ] Cloninger et al., [ 26,27 ] Lecommandoux et al., [ 28 ] Elling and co‐workers, [ 29–31 ] Percec et al., [ 32–35 ] Putnam et al., [ 36 ] and more recently Rosencrantz et al. [ 37 ] This approach takes advantage of the ability of galectin‐3 to oligomerize in the presence of multivalent ligands leading to an effective increase in binding avidity.…”

“…[ 37 ] This approach takes advantage of the ability of galectin‐3 to oligomerize in the presence of multivalent ligands leading to an effective increase in binding avidity. [ 18–35,38 ]…”

High-Sensitivity Detection of the Lung Cancer Biomarker CYFRA21-1 in Serum Samples Using a Carboxyl-MoS2 Functional Film for SPR-Based Immunosensors

“…The results showed that the angular shift caused by the traditional SPR (MOA-based) chip was similar at flow rates of 30 and 60 µL/min. We performed a 1:1 interaction of the SPR biosensor sensing surface with anti-CYFRA21-1 protein using the binding Langmuir model (Bumba et al, 2018). The kinetic analysis for the traditional SPR chip at a flow rate of 60 µL/min yielded an affinity binding constant (K A ) of 1.78 × 10 8 M −1 , association rate constant (k a ) of 2.71 × 10 4 M −1 S −1 and dissociation rate constant (k d ) of 15.23 × 10 −5 S −1 .…”

Core-2O-glycans are required for galectin-3 interaction with the osteoarthritis related protein lubricin

“…Previous studies have indicated that galectin-3 exhibits only weak affinity to O-glycoproteins and O-glycopeptides expressing single core 1 O-glycans (Galb1-3GalNAc or NeuAcα2-3Galb1-3GalNAc) 34,42 . Instead, internal LacNAc glycans on extended oligosaccharides have been described to be the high affinity ligands for galectin-3 43 . Here, we suggest that interaction between galectin-3 and native lubricin is mediated by short single LacNAc extended core 2 structures.…”