Publication details
Calocera viscosa lectin – a fungi member of the jacalin-related lectins family
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| Year of publication | 2023 |
| Type | Appeared in Conference without Proceedings |
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| Description | The research is focused on the properties and structural analysis of the new lectin – saccharide binding protein – from the mushroom Calocera viscosa (CalVL). Mushroom lectins have been variously explored and studied for their potential in biomedicine and diagnostics. Based on their interactions with glycans, they exhibit mitogenic, antimicrobial, and antiproliferative effects.1 For the characterization of this lectin’s properties, various methods were employed. Inceptive predictions suggested that the CalVL, with its ß-prism I fold, should belong to the family of jacalin related lectins (JRLs). JRLs may be briefly divided based on their preference of binding to D galactose and D-mannose.2 Molecular cloning and tests of gene expression were performed, and the CalVL was produced in the Escherichia coli expression system. Later, it was purified using affinity chromatography on mannose-agarose resin. Later it was shown by agglutination assays that the CalVL is able to agglutinate both yeast cells and human erythrocytes due to the interaction with their surface saccharides. Further analysis of the binding properties was inspected by the Glycan Array. Thermostability of the CalVL was determined by Differential Scanning Fluorimetry, homogeneity, and the oligomeric state of the protein by Dynamic Light Scattering and Analytical Ultracentrifugation. The CalVL’s ability to crystallize was tested in various screens using the vapor diffusion method, specifically the sitting drop technique. Data collection from obtained crystals was performed on synchrotron PETRA III and BESSY II in Hamburg and Berlin, respectively. The structure of the CalVL is in the process of solving but preliminary, there is a CalVL without any ligand, and with D mannose. The phase problem of the measured data was solved by molecular replacement, where the model of the CalVL predicted by the AlphaFold2 was utilized. These structures confirm the prediction of the CalVL being a member of the JRLs family. |
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