Loss of glycosylation affects binding of Anthrax toxin to TEM8 but not to CMG2.
A and C) HeLa cells were transfected for 48h with the respective cDNAs. Cells were treated for 1h at 4°C with 500 ng/ml PA83 and shifted to 37°C for 10 min to induce cleavage and heptamerization. Immunoprecipitates against TEM8-HA/CMG2-V5 were analyzed by SDS-PAGE and Western Blotting against PA and TEM8-HA/CMG2-V5. Control cells are non-transfected. D50A is a binding deficient mutant that serves as a negative control. B and D) HeLa cells were transfected for 48h with the respective cDNAs. Cells were lysed and incubated for 1h at 4°C with 1 μg/ml PA83. Immunoprecipitates against TEM8-HA/CMG2-V5 were analyzed by SDS-PAGE and Western Blotting against PA and TEM8-HA/CMG2-V5. Control cells are non-transfected. D50A is a binding deficient mutant that serves as a negative control.
