Isolation and characterization of a lectin from Grifola frondosa fruiting bodies

An N-acetylgalactosamine-specific lectin (GFL) was isolated from Grifola frondosa fruiting bodies by affinity chromatographies on acid-treated Sepharose CL-4B and then GalNAc-Toyopearl. The isolated lectin agglutinated all types of erythrocytes equally. Molecular masses estimated by gel filtration under various buffers and matrices varied from 30 to 52 kDa. On the other hand, SDS-PAGE in the presence or absence of 2-mercaptoethanol showed three major bands of 33, 66 and 100 kDa and a faint band of 65 kDa. This lectin exhibited GalNAc-specificity. The protein was a glycoprotein containing 3.3% total sugar, and the amino acid analysis revealed a high content of acidic and hydroxy amino acids and a low content of methionine and histidine. GFL was cytotoxic against HeLa cells. The toxicity did not appear after preincubating the lectin with the haptenic sugar N-acetylgalactosamine.     

The wound-healing responses of Antithamnion nipponicum and Griffithsia pacifica (Ceramiales, Rhodophyta) monitored by lectins

The binding of FITC labeled lectins to repair cells of Antithamnion nipponicum Yamada et Inagaki and Griffithsia pacifica Kylin, and their physiological effects on somatic cell fusion have been studied. Results indicate that repair cells strongly bind the lectins ConA and LCA, whereas other lectins did not bind to the cell, The binding of these lectins to the dead cell wall shows ConA and LCA specific substances are secreted from the tip of the repair cells. When fluorescently labeled ConA or LCA was added at various time intervals after wounding, it firstly bound (3 h post-wounding) as a thin layer at the tips of the adjacent cells. Later (4–5 h post-wounding) labeling also appeared at the tips of the repair ceils. Intense labeling at these sites continued throughout the wound-healing process until repair cell fusion, at which time the lectin labeling was reduced to a narrow ring around the area of fusion, When added to plants prior to wounding and with continued monitoring, these same lectins were found to act as inhibitors to the wound-healing response. Other control lectins showed no inhibitory effects. These results suggest that a signal glycoprotein with α-D-mannosyl residues is involved in the wound-healing process of Antithamnion nipponicum. Lectins conjugated with visible tags can be used as a very fast and useful tool to monitor these signal substances.