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Updated: May 21, 2026
Rapid Glyco-Qualitative Assessment of Recombinant Proteins Using a Fully Automated System
Published on: June 28, 2024
Characterization and cloning of GNA-like lectin from the mushroom Marasmius oreades
Michiko Shimokawa1, Ayako Fukudome, Ryoko Yamashita
1Department of Applied Biological Chemistry, The United Graduate School of Agricultural Sciences, Kagoshima University, 1-21-24 Korimoto, Kagoshima 890-0065, Japan.
Abstract:
A new mannose-recognizing lectin (MOL) was purified on an asialofetuin-column from fruiting bodies of Marasmius oreades grown in Japan. The lectin (MOA) from the fruiting bodies of the same fungi is well known to be a ribosome-inactivating type lectin that recognizes blood-group B sugar. However, in our preliminary investigation, MOA was not found in Japanese fruiting bodies of M. oreades, and instead, MOL was isolated. Gel filtration showed MOL is a homodimer noncovalently associated with two subunits of 13 kDa. The N-terminal sequence of MOL was blocked. The sequence of MOL was determined by cloning from cDNA and by protein sequencing of enzyme-digested peptides. The sequence shows mannose-binding motifs of bulb-type mannose-binding lectins from plants, and similarity to the sequences. Analyses of sugar-binding specificity by hemagglutination inhibition revealed the preference of MOL toward mannose and thyroglobulin, but asialofetuin was the strongest inhibitor of glycoproteins tested. Furthermore, glycan-array analysis showed that the specificity pattern of MOL was different from those of typical mannose-specific lectins. MOL preferred complex-type N-glycans rather than high-mannose N-glycans.
Insights
A novel mannose-recognizing lectin (MOL) was isolated from Marasmius oreades mushrooms. This lectin displays unique binding preferences for complex-N-glycans, differing from typical mannose-specific lectins.
Area of Science:
- Biochemistry
- Molecular Biology
- Glycobiology
Background:
- Marasmius oreades mushrooms are known to contain lectins, such as MOA, which recognize blood-group B sugar.
- Previous studies on M. oreades lectins primarily focused on MOA, a ribosome-inactivating lectin.
- A discrepancy was noted in Japanese M. oreades fruiting bodies, where MOA was absent.
Purpose of the Study:
- To isolate and characterize a novel mannose-recognizing lectin (MOL) from Japanese Marasmius oreades fruiting bodies.
- To determine the structural and functional properties of the newly identified lectin.
- To elucidate the specificities of MOL's carbohydrate-binding activity.
Main Methods:
- Purification of MOL using an asialofetuin-affinity column.
- Gel filtration chromatography to determine molecular weight and subunit association.
- N-terminal sequencing, cDNA cloning, and protein sequencing of enzyme-digested peptides for sequence determination.
- Hemagglutination inhibition assays and glycan-array analysis to assess sugar-binding specificity.
Main Results:
- A novel mannose-recognizing lectin (MOL) was successfully isolated from Japanese M. oreades.
- MOL is a homodimer composed of two 13 kDa subunits, with a blocked N-terminal sequence.
- Sequence analysis revealed mannose-binding motifs characteristic of plant bulb-type lectins.
- MOL demonstrated a preference for mannose and thyroglobulin, with asialofetuin as the strongest glycoprotein inhibitor.
- Glycan-array analysis indicated that MOL preferentially binds complex-type N-glycans over high-mannose N-glycans, distinguishing it from typical mannose-specific lectins.
Conclusions:
- A new mannose-binding lectin (MOL) has been identified in Japanese Marasmius oreades.
- MOL possesses distinct carbohydrate-binding specificities, favoring complex N-glycans.
- The findings expand the understanding of lectin diversity within the Marasmius genus and their glycan recognition capabilities.
