Swamy Bale M, Bhat Anuradha G, Hegde Ganapati V, Naik Ramachandra S, Kulkarni Srikanth, Inamdar Shashikala R
Department of Biochemistry, Karnatak University, Dharwad 580 003, India.
Glycobiology. 2004 Nov;14(11):951-7. doi: 10.1093/glycob/cwh130. Epub 2004 Jul 14.
Many fungi are known to secrete lectins, but their functional roles are not clearly understood. Sclerotium rolfsii, a soilborne plant pathogenic fungus capable of forming fruiting bodies called sclerotial bodies, secrete a cell wall-associated Thomsen-Friedenreich antigen-specific lectin. To understand the functional role of this lectin, we examined its occurrence and expression during development of the fungus. Furthermore, putative endogenous receptors of the lectin were examined to substantiate the functional role of the lectin. Immunolocalization studies using FITC-labeled lectin antibodies revealed discrete distribution of lectin sites at the branching points of the developing mycelia and uniformly occurring lectin sites on the mature sclerotial bodies. During development of the fungus the lectin is expressed in small amounts on the vegetative mycelia and reaching very high levels in mature sclerotial bodies with a sudden spurt in secretion at the maturation stage. Capping of the lectin sites on the sclerotial bodies by lectin antibodies or haptens inhibit strongly the germination of these bodies, indicating functional significance of the lectin. At the maturation stage the lectin interacts with the cell wall-associated putative endogenous receptor leading to the aggregation of mycelium to form sclerotial bodies. The lectin-receptor complex probably acts as signaling molecule in the germination process of sclerotial bodies. Using biotinylated lectin, the receptors were identified by determining the specific lectin binding to lipid components, extracted from sclerotial bodies, and separated on thin-layer chromatograms. Preliminary characterization studies indicated that the receptors are glycosphingolipids and resemble inositolphosphoceramides. These findings together demonstrate the importance of lectin-receptor interactions to explain hitherto speculated functional role of the lectins and also the glycosphingolipids of fungi.
已知许多真菌会分泌凝集素,但其功能作用尚不清楚。齐整小核菌是一种能形成称为菌核体的子实体的土传植物病原真菌,它会分泌一种与细胞壁相关的、针对桑福德 - 弗里德赖希抗原的特异性凝集素。为了解这种凝集素的功能作用,我们研究了其在真菌发育过程中的出现情况和表达情况。此外,还对该凝集素的假定内源性受体进行了研究,以证实该凝集素的功能作用。使用异硫氰酸荧光素标记的凝集素抗体进行的免疫定位研究表明,凝集素位点在发育中的菌丝体分支点处呈离散分布,而在成熟菌核体上则均匀分布。在真菌发育过程中,凝集素在营养菌丝体上少量表达,在成熟菌核体中达到非常高的水平,在成熟阶段分泌量会突然激增。用凝集素抗体或半抗原封闭菌核体上的凝集素位点会强烈抑制这些菌核体的萌发,这表明凝集素具有功能意义。在成熟阶段,凝集素与细胞壁相关的假定内源性受体相互作用,导致菌丝体聚集形成菌核体。凝集素 - 受体复合物可能在菌核体的萌发过程中作为信号分子起作用。使用生物素化凝集素,通过测定从菌核体中提取并在薄层色谱图上分离的脂质成分与特异性凝集素的结合来鉴定受体。初步表征研究表明,这些受体是糖鞘脂,类似于肌醇磷酸神经酰胺。这些发现共同证明了凝集素 - 受体相互作用对于解释迄今推测的凝集素功能作用以及真菌糖鞘脂的重要性。