Figure 2.

Role of ribosome and translocon complex on nascent chain folding. BHK cells infected by recombinant SFV virus expressing HA-RAP were labeled for 1 min and chased in various conditions. The lysates were immunoprecipitated by α-NHA and analyzed by 5.5% two-dimensional gels. The markers are wild-type HA (80 kDa), muscle phosphorylase (97 kDa). and Escherichia coli β-galactosidase (115 kDa). (A) One-minute pulse for HA-RAP. (B) One-minute pulse followed by 6-min chase with 1 mM cysteine and 1 mM methionine. Shown is a very short exposure to demonstrate clear separation between IT1 and IT2. (C) One-minute pulse followed by 20-min chase in the presence of 1 mM puromycin. (D) One-minute pulse followed by 20-min chase in the presence of 1 mM cycloheximide.