An endogenous carbohydrate-binding agglutinin of BHK cells. Purification, specificity and interaction with normal and ricin-resistant cell lines.
Several lectin-like activities were detected on the surface of unfixed, viable BHK cells by reaction with FITC-labelled glycosylated albumin derivatives. A prominent surface staining was obtained with the beta-lactosyl, beta-N-acetylgalactosaminyl, alpha-mannosyl and beta-N-acetylglucosaminyl derivatives. Endogenous lectin-like activities were also detected in BHK cell homogenates by haemagglutination of glutaraldehyde-fixed rabbit erythrocytes. Haemagglutinating activity was purified by chromatography of sodium deoxycholate-extracts of BHK cell microsomal fractions on Sepharose 4B and asialofetuin-Sepharose 4B. A purified agglutinin was eluted from the latter column with 0.2 M thiodigalactoside. The haemagglutination mediated by the purified factor was inhibited by thiodigalactoside, N-acetylgalactosamine, galactosyl-beta 1-4-N-acetylglucosamine and several glycoproteins. The purified agglutinin agglutinated trypsinised, fixed normal BHK cells more readily than several ricin-resistant cell lines. By contrast, a mannose-binding lectin from rabbit serum reacted equally well with normal and mutant cells. These results are in general agreement with models of cell-cell adhesion involving the interaction of surface located lectins with carbohydrate sequences of normal BHK cell surface glycoproteins.
D000818:Animals; D002241:Carbohydrates; D002460:Cell Line; D002462:Cell Membrane / Q000032:analysis / Q000648:ultrastructure; D006224:Cricetinae; D004351:Drug Resistance; D016650:Fluorescein-5-isothiocyanate; D005452:Fluoresceins; D005456:Fluorescent Dyes; D006384:Hemagglutination; D007668:Kidney; D037102:Lectins / Q000302:isolation & purification*; D011817:Rabbits; D012276:Ricin / Q000633:toxicity*; D013347:Subcellular Fractions / Q000032:analysis; D013861:Thiocyanates
D002241:Carbohydrates; D005452:Fluoresceins; D005456:Fluorescent Dyes; D037102:Lectins; D013861:Thiocyanates; D012276:Ricin; D016650:Fluorescein-5-isothiocyanate