Concanavalin A binding induces association of possible mating-type receptors with the cytoskeleton in Tetrahymena.
The lectin concanavalin A (conA; 25 micrograms/ml) inhibits conjugation in the ciliate Tetrahymena, and binds to receptors localized at the junction between conjugating cells. We report here that succinyl-conA (30 micrograms/ml) has similar activity, but that two other mannosespecific lectins, lentil and pea lectins, have inhibitory activities more than tenfold lower in this system, indicating that factors other than mannose specificity are essential for biological activity. By using fluorescein-isothiocyanate (FITC)-conA, we have found that extraction of cells with the detergent Triton X-100 removes conA receptors from the extraction-resistant cytoskeleton, but that the binding of conA to its receptor before extraction associates the ligand-receptor complex with the cytoskeleton. Under the hypothesis that the conA receptor may be a mating type receptor, we have used this ligand-induced differential cytoskeletal association, in conjunction with electrophoresis and Western blotting, to identify a glycoprotein with an apparent molecular weight (MW) of 23,000 D which may be a mating type receptor. Our data are consistent with a model in which a direct interaction between the conA receptor and the cytoskeleton, rather than receptor cross-linking, is the biologically significant activity of ligand binding.
D000818:Animals; D003208:Concanavalin A / Q000031:analogs & derivatives / Q000378:metabolism / Q000494:pharmacology*; D003599:Cytoskeleton / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D016650:Fluorescein-5-isothiocyanate; D005452:Fluoresceins; D005456:Fluorescent Dyes; D007159:Immunologic Tests; D008856:Microscopy, Fluorescence; D011952:Receptors, Concanavalin A / Q000378:metabolism*; D012098:Reproduction; D013768:Tetrahymena / Q000502:physiology* / Q000648:ultrastructure; D013861:Thiocyanates
D005452:Fluoresceins; D005456:Fluorescent Dyes; D011952:Receptors, Concanavalin A; D013861:Thiocyanates; C027774:concanavalin A-binding glycoproteins; D003208:Concanavalin A; C019558:succinylconcanavalin A; D016650:Fluorescein-5-isothiocyanate