Structure of Galectin-3 bound to a model membrane containing ganglioside GM1

Galectin-3 (Gal-3) is a 0-galactosidase-binding protein involved in various biological processes, including neuronal growth and adhesion. The pairing of Gal-3 with ganglioside GM1's pentasaccharide chain at the outer leaflet of the plasma membrane, which triggers downstream cell-signaling cascades, seems to be involved in these processes. A crucial feature of Gal-3 is its ability to form oligomers and supramolecular assemblies that connect various carbohydrate-decorated mol-ecules. Although we know the atomistic structure of Gal-3 bound to small carbohydrate ligands, it remains unclear how Gal-3 binds GM1 in a membrane. Furthermore, the influence of this interaction on Gal-3's structure and oligomeric assembly has to be elucidated. In this study, we used X-ray reflectivity (XR) from a model membrane to determine the structure and surface coverage of Gal-3 bound to a membrane containing GM1. We observed that the carbohydrate recognition domain interacts with GM1's pentasaccharide, while the N-terminal domain is pointed away from the membrane, likely to facilitate protein-protein in-teractions. In a membrane containing 20 mol % GM1, Gal-3 covered-50% of the membrane surface with one Gal-3 molecule bound per 2130 A2. We used molecular dynamics simulations and Voronoi tessellation algorithms to build an atomistic model of membrane-bound Gal-3, which is supported by the XR results. Overall, this work provides structural information describing how Gal-3 can bind GM1's pentasaccharide chain, a prerequisite for triggering regulatory processes in neuronal growth and adhesion.

Automated summary

In this study, X-ray reflectivity was used to determine the structure and surface coverage of Gal-3 bound to a model membrane containing GM1. The carbohydrate recognition domain of Gal-3 interacted with GM1's pentasaccharide chain, while the N-terminal domain was pointed away from the membrane. This work provides important structural information on the interaction between Gal-3 and GM1.