Figure 2. PRDC^WT and PRDC^C120S are dimeric.

A) Western blot analysis of PRDC^WT produced in E. coli (lanes 1 and 3) and in HEK293F (lanes 2, 4-6). If indicated, samples were treated with 5% 2-mercaptoethanol (BME) to reduce disulfide bonds prior to gel loading. PRDC^WT produced in HEK293F cells was purified by His affinity resin (lane 5) and deglycosylated with PNGaseF (lane 6). B) Western blot analysis of PRDC^C120S produced in E. coil and HEK293F similar to (A). A polyclonal anti-PRDC antibody was used for detection in (A) and (B). Size exclusion elution profile of PRDC^WT, PRDC^C120S and molecular weight standards. Purified proteins (100 μg) were applied to a Superdex 75 column. D) Sedimentation coefficient c(s) distribution profile of PRDC^WT and PRDC^C120S (1 mg/ml) determined by sedimentation velocity. After fitting for the frictional ratio (f/f₀) the c(s) distribution was transformed into a c(M) distribution (not shown) to determine the molecular mass estimates (labeled).