Conformational fingerprint of mutant ACE.

<p><b>A–D.</b> Precipitation of ACE activity from cell lysates (membrane-bound form) and culture medium (soluble form) from HEK and CHO cells expressing WT and mutant (Y465D) ACE was estimated using 16 mAbs to the epitopes localized on the N and C domains of human ACE. Samples were equilibrated according to 5 mU/ml of the ACE activity with Hip-His-Leu as a substrate and incubated with a wells on the microtiter plate covered by mAbs to ACE via goat-anti-mouse IgG; then precipitated ACE activity was quantified by fluorimetric assay - plate precipitation assay <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0025952#pone.0025952-Danilov4" target="_blank">[42]</a>. Data are expressed as a ratio of ACE activity precipitated by different mAbs from mutant ACE to that of WT ACE. Data presented as a mean of 3–4 independent determinations in duplicate. * p<0.05 indicates ratio shown is significantly different from 1. mAbs showing significant changes in binding compared to WT are colored (red: increase >20%, yellow: decrease >20%).