ナマズ卵レクチンのクラスター形成と細胞内輸送

Rhamnose-binding lectins are widely found in fush eggs,and Silurus asotus lectin (SAL) isolated from catfish eggs having three carbohydrate recognition domains preferentially recognizes non-reducing end Galα-linked sugar chain.In the previous study,we revealed that SRL binds to globotriaosylceramide (Gb3) by the surface plasmon resonance analysis.However,its biological effect on cultured cells is still unclear.To investigate localization and trafficking of SAL in the renel adenocarcinoma ACHN cells,which express Gb3 on the cell surface,we prepared HiLyte Fluor 555-labeled SAL (HL-SAL).When ACHN cells were treated with HL-SAL at 4℃ for 5min, and at 37℃ for 24h,HL-SAL was distributed on the cell membrane and in the intracellular compartment,respectively.To trace the trafficking route of HL-SAL from cell surface to the intracellular compartment,the images of HL-SAL-treated live cells were obtained using confocal scanning microscopy.HL-SAL was clustered on ACHN cell surface,and furthermore,partially co-localised with transferrin in intracellular compartment.These results suggest that SAL induces alteration of Gb3 distribution on the membrane and migrates from the cell surface to the intracellular vesicles