Differential subcellular localization of wild type and mutant ACE.

Confocal microscopy was used to determine the localization of WT- vs. Q1069R-ACE mutant expressed in CHO cells following immunostaining with anti-ACE mAb 9B9 and Alexa 488 goat anti-mouse secondary Ab. A-C: WT-ACE expressing cells; D-F: Q1069R-ACE expressing cells. A,B and D,E. Immunostaining of WT and Q1069R ACE in 0.3% paraformaldehye for 15 min at 4C (fixed only). B and E. Treatment with the chemical chaperone sodium butyrate and proteosome inhibitor MG132 for 24 hrs followed by immunostaining with 9B9 mAb. C and F. CHO cells fixed and permeabilized with 3% paraformaldehyde for 15 min at 37C and labeled as above show WT-ACE on the plasma membrane and to a lesser extent in the cytosol and Q1069R-ACE in perinuclear areas.