Crystal structure of MOA in complex with a peptide fragment: A protease caught in flagranti

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 elucidate the inhibitory roles of zinc and cadmium towards papain-like cysteine proteases in general. The reported structures, together with activity data of MOA active site variants, point to a conservation of the general proteolysis mechanism established for papain. Ultimately, the findings suggest that papain and the papain-like domain of MOA are the product of convergent evolution.
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Highlights • MOA is a chimerolectin with protease activity carrying a unique papain-like fold. • The structures of inactive MOA variants reveal cleaved peptides in the active site. • Anomalous diffraction data elucidate activation or inhibition by different metals. • The MOA papain-like domain is a product of convergent evolution. • MOA is the holotype of an emerging family of enzymatically active fungal lectins.