TY - JOUR
T1 - Angiotensin-converting enzyme 2 catalytic activity in human plasma is masked by an endogenous inhibitor
AU - Lew, Rebecca Ann
AU - Warner, Fiona Jane
AU - Hanchapola, Iresha
AU - Yarski, Michael Anthony
AU - Manohar, Jay
AU - Burrell, Louise M
AU - Smith, Alexander Ian
PY - 2008
Y1 - 2008
N2 - Angiotensin converting enzyme-2 (ACE2) is thought to act in an opposing manner to its homologue, angiotensin converting enzyme (ACE), by inactivating the vasoconstrictor peptide angiotensin II and generating the vasodilatory fragment, angiotensin 1-7. Both ACE and ACE2 are membrane-bound ectoenzymes and may circulate in plasma as a consequence of a proteolytic shedding event. In this study, we show that ACE2 circulates in human plasma, but its activity is suppressed by the presence of an endogenous inhibitor. Partial purification of this inhibitor indicated that the inhibitor is small, hydrophilic, and cationic, but not a divalent metal cation. These observations led us to develop a method for removal of the inhibitor, thus allowing detection of plasma ACE2 levels using a sensitive quenched fluorescent substrate-based assay. Using this technique, ACE2 activity measured in plasma from healthy volunteers (n = 18) ranged from 1.31 to 8.69 pmoles substrate cleaved/min/mL (mean +/- S.E.M. = 4.44 +/- 0.56 pmol/min/mL). Future studies of patients with cardiovascular, renal and liver disease will determine whether plasma ACE2 is elevated in parallel with increased tissue levels observed in these conditions.
AB - Angiotensin converting enzyme-2 (ACE2) is thought to act in an opposing manner to its homologue, angiotensin converting enzyme (ACE), by inactivating the vasoconstrictor peptide angiotensin II and generating the vasodilatory fragment, angiotensin 1-7. Both ACE and ACE2 are membrane-bound ectoenzymes and may circulate in plasma as a consequence of a proteolytic shedding event. In this study, we show that ACE2 circulates in human plasma, but its activity is suppressed by the presence of an endogenous inhibitor. Partial purification of this inhibitor indicated that the inhibitor is small, hydrophilic, and cationic, but not a divalent metal cation. These observations led us to develop a method for removal of the inhibitor, thus allowing detection of plasma ACE2 levels using a sensitive quenched fluorescent substrate-based assay. Using this technique, ACE2 activity measured in plasma from healthy volunteers (n = 18) ranged from 1.31 to 8.69 pmoles substrate cleaved/min/mL (mean +/- S.E.M. = 4.44 +/- 0.56 pmol/min/mL). Future studies of patients with cardiovascular, renal and liver disease will determine whether plasma ACE2 is elevated in parallel with increased tissue levels observed in these conditions.
UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=18223027
M3 - Article
SN - 0958-0670
VL - 93
SP - 685
EP - 693
JO - Experimental Physiology
JF - Experimental Physiology
IS - 5
ER -