Effect of ACE mutations on blood ACE phenotype parameters

Figures

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  Fig 1. Localization of tested ACE mutations in the N and C domains of ACE. The positions of ACE mutations are shown on the Cryo-EM structure of the truncated (1–1201) human somatic ACE (PDB 7Q3Y) [18] using molecular surface representation. Key amino acids are denoted using somatic ACE numbering. The surface is colored light beige with specific amino acid residues color coded as follows: ACE mutations are highlighted in magenta and additionally marked by arrows; putative Asn glycosylation sites are highlighted in green; the first residue in the N-domain of somatic ACE is marked with its number L1, while the last visible residue in the C-terminal end of this truncated somatic ACE is marked with its number S1201. Both terminal residues are highlighted in blue. The epitopes for some mAbs to the N and C domains are shown as black circles for orientation with a diameter 30 Å, which corresponds to 700 Å2 of the area covered by this mAb. Insert: the mutations I989T, V992M and V997M are highlighted in magenta; active site residues are highlighted in red.

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  Fig 2. Quantification of blood ACE levels in carriers of ACE mutations. Blood ACE protein was precipitated from EDTA-plasma by mAb 9B9 (with epitope on the surface of the N domain of ACE), and its activity was quantified fluorometrically using ZPHL as a substrate. A) The immunoreactive ACE protein was quantified in plasma samples of 15 newborn carriers with 11 different ACE mutations. The asterisk indicates that ACE in the blood of patient VKK056 had two mutations, T887M and Y215C. B) ACE levels in plasma samples were adjusted according to the donor’s genotype I/D polymorphism [25, 26]. ACE levels in the blood of each person were presented as the mean of several experiments with standard deviation. C) Predictions of the potential damaging effects of 11 mutations on the ACE protein using 4 different predictive tools were derived from S1 Table in S1 File [1]. Values shown in red are predicted to be damaging by the listed predictive engine; values in black are predicted to be benign. Data were expressed as % of ACE levels from the corresponding value for control pooled plasma samples from donors without ACE mutations (green bars). Bars highlighted with orange and brown indicate samples…

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  Fig 3. ACE phenotype for carriers of ACE mutations. A-B. Blood ACE protein was precipitated by mAb to the N domain (9B9), and precipitated ACE activity was quantified with substrate ZPHL (A) and HHL as in Fig 2. The ZPHL/HHL ratio was calculated for each carrier of the different ACE mutations (B). C) Precipitation of ACE protein from the blood of carriers of some ACE mutations using mAbs with various epitopes, including the region of these mutations normalized to the precipitation of ACE from the same sample by mAb 9B9. The coloring is the same as in Fig 2.

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  Fig 4. The position of the mutations I989T, V992M and V997M in the ACE C domain. The positions of ACE mutations are shown on the Cryo-EM structure of the truncated (1–1201) human somatic two-domain ACE (PDB 7Q3Y) [18] using ribbon representation. The coloring is the same as in Fig 1.