Fig. 1

Targeted PAM proteins. a. Diagram of PAM1 and GFP-tagged targeted PAM proteins. The regions of ERGIC-53, Galactosyl Transferase 1, cation independent mannose-6-phosphate receptor and LAMP1 used to redirect rat PAM/GFP are indicated. PAM820s/GFP was truncated immediately after the catalytic core of PAL. b. pEAK Rapid cells, a derivative of hEK293 cells, were transiently transfected with vectors encoding the indicated proteins. After 24 h, cell extracts were prepared using 20 mM NaTES, 10 mM mannitol, 1 % TX-100, pH 7.4; aliquots were subjected to Western blot analysis and visualized using an antibody to the PHM domain. c. PHM activity was measured in each cell lysate and normalized to the amount of PHM protein detected on the Western blot; the PHM activity/PHM protein ratio for PAM/GFP was set to 1.0 for each of three experiments; error bars show the standard deviation. None of the differences were significant