| Products/Services Used | Details | Operation |
|---|---|---|
| Peptide Synthesis | MOA crystals grew from a solution containing the purified protein at a concentration of 5 mg/ml and the trisaccharide Galα1,3(Fucα1,2)Gal (Dextra; 1:20 MOA:sugar molar ratio), pre-mixed directly before the experiments were set up. Hanging-drop vapor diffusion experiments were carried out using 24-well tissue culture plates (TPP, Sigma-Aldrich) and siliconized glass cover slides (Hampton Research), on which 1 μl crystallization solution was added to an equal volume of the protein-ligand solution. Crystals of MOA in complex with peptide substrates PVPRAHS or PVVRAHS (GenScript, USA) were grown according to the same protocol detailed above, pre-mixing sugar and protein, and additionally adding one of the peptides at a 1:10 MOA:peptide molar ratio. Details of the crystallization conditions are reported in Table S1. Rod-shaped crystals grew over the course of 2-4 weeks at 20°C, with dimensions of 0.05 mm × 0.05 mm × 0.4 mm. | Get A Quote |
The Marasmius oreades agglutinin (MOA) is the holotype of an emerging family of fungal chimerolectins and an active Ca2+/Mn2+-dependent protease, which exhibits a unique papain-like fold with special active site features. Here we investigated the functional significance of the structural elements differentiating MOA from other papain-like cysteine proteases. X-ray crystal structures of MOA co-crystallized with two synthetic substrates reveal cleaved peptides bound to the catalytic site, corresponding to the final products of the proteolytic reaction. Anomalous diffraction data on crystals grown in the presence of calcium and manganese, cadmium or zinc resolve the calcium/manganese preference of MOA and elucid... More
