Figure 2.
The C-terminal half of the D domain is necessary for intracellular retention of Dd-PDI. (A) A stable clonal cell line was created and shown to express about threefold more Dd-PDIΔC (●) than the endogenous Dd-PDI (▴), as revealed by anti-PDI immunoblotting. This cell line was then investigated for secretion/retention of Dd-PDI and recombinant Dd-PDIΔC by pulse/chase labeling with [35S]Met followed by anti-PDI immunoprecipitation. At the indicated times, aliquots were retrieved and cells (B, intracellular) were separated from medium (C, extracellular) by centrifugation and analyzed separately. After quantification by phosphorimager, the normalized signals corresponding to full-length Dd-PDI (▴) and Dd-PDIΔC (●) were plotted as a function of time.