Figure 7.
Pulse–chase analysis of HA-tagged Kex2p and CPY. Cells transformed with the indicated plasmids were labeled for 10 min at 30°C with Tran35S label, and a chase was begun. Samples were harvested at the indicated times and subjected to immunoprecipitation. (A) HA-tagged proteins were immunoprecipitated with the monoclonal 12CA5 anti-HA antibody. (B) Data from three independent experiments were digitally scanned. Values are expressed as percentage of the maximum intensity in each experiment, and the log of the value was used for linear regression analysis to determine the half-life of Kex2p protein in a strain transformed with the KEX2HA construct (squares), Δprokex2HA construct (circles), or Δprokex2HA + preprokex2 constructs (triangles). (C) Anti-CPY antibodies immunoprecipitated ER and Golgi forms of pro-CPY (p1 and p2, respectively) and mature CPY (m).