Figure 1

AMDM mutants are not processed to the fully glycosylated and phosphorylated form of GC-B. WT and the indicated missense versions of GC-B were isolated from transiently transfected 293T cells by sequential immunoprecipitatin/SDS-PAGE purification. The resulting gel was incubated with ProQ Diamond dye to determine GC-B phosphate levels (lower panel). The same gel was then washing and stained with Coomassie to determine the amount of GC-B protein present (upper panel). MW in Kilo-Daltons is shown for protein standards on left. Abbreviations are: GFP, green fluroscent protein; 7A, GC-B containing alanine substitutions for all seven known phosphorylation sites.