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. 2009 May 1;297(1):L84–L96. doi: 10.1152/ajplung.00071.2009

Fig. 5.

Fig. 5.

ACE2 mutant ACE2-L584A attenuates sACE2 release from cells. A: diagram of ACE2-L584A ACE2 mutants and chimeric ACE2-CD4/TM-Cyto (see Fig. 5A). B: immunodetection of cell-associated and sACE2. Results indicated that mutants of ACE2 (ACE2-L584A) and ACE2-CD4/TM-Cyto (ACE2-CD4/TM-Cyto/L584A) were not released from respective transfected HEK293 cells. C: chromatographs of ANG II to ANG (1–7) conversion by cell culture media collected from cells expressing either wild-type ACE2 or ACE2-L584A protein. Left: ACE2 transfected HEK293 cell culture medium efficiently converted ANG II to ANG (1–7) and PMA further enhanced the conversion efficiency. Right: culture medium from ACE2-L584A mutant shows no conversion of ANG II to ANG (1–7). PMA-stimulated ACE2-L584A transfected cell culture medium also showed no converting activity, consistent with an absence of shedding.