Active Angiotensin I Converting Enzyme 2 (ACE2)

ACEH; ACEII; ACE-II; Peptidyl-Dipeptidase A; ACE-related carboxypeptidase; Angiotensin-converting enzyme homolog; Metalloprotease MPROT15

ACTIVITY TEST

The activity of recombinant human ACE2 is also measured by its ability to cleave a fluorogenic peptide substrate MCA-Tyr-Val-Ala-Asp-Ala-Pro-Lys(DNP)-OH in the assay buffer 50 mM Tris, 1 M NaCl, pH 7.5. The rhACE2 is diluted to 0.5 ug/mL in assay buffer. Loading into a black well plate 50 µL of 0.5 ug/mL rhACE2 and start the reaction by adding 50 µL of 20 µM substrate, with a substrate blank containing 50 µL assay buffer, 50 µL substrate, and no rhACE2. Then read at excitiation and emission wavelengths of 320 nm and 405 nm, respectively, in kinetic mode for 5 minutes. The specific activity of recombinant human ACE2 is > 1100 pmol/min/µg.

Angiotensin-converting enzyme 2 (ACE2), as a transmembrane protein, serves as the main entry
point into cells for some coronaviruses, including HCoV-NL63, SARS-CoV , and SARS-CoV-2 . More specifically, the binding of the spike S1 protein of SARS-CoV and SARS-CoV-2 to the enzymatic domain of ACE2 on the surface of cells results in endocytosis and translocation of both the virusand the enzyme into endosomes located within cells. Besides, recent studies show that spike (S) proteins of 2019-nCoV and SARS-CoV may use the same host cell receptor called angiotensin-converting enzyme 2 (ACE2) for entering into host cells, thus a binding ELISA assay was conducted to detect the interaction of recombinant human ACE2 and recombinant Spike glycoprotein RBD. Briefly, biotin-linked ACE2 were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100μl were then transferred to RBD-coated microtiter wells and incubated for 2h at 37℃. Wells were washed with PBST 3 times and incubation with HRP conjugage for 30min, then wells were aspirated and washed 5 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37℃. Finally, add 50µl stop solution to the wells and read at 450nm immediately. The binding activity of ACE2 and RBD was shown in Figure 1, and this effect was in a dose dependent manner.

USAGE

Reconstitute in 10mM PBS (pH7.4) to a concentration of 0.1-1.0 mg/mL. Do not vortex.

STORAGE

Avoid repeated freeze/thaw cycles. Store at 2-8°C for one month. Aliquot and store at -80°C for 12 months.

STABILITY

The thermal stability is described by the loss rate. The loss rate was determined by accelerated thermal degradation test, that is, incubate the protein at 37°C for 48h, and no obvious degradation and precipitation were observed. The loss rate is less than 5% within the expiration date under appropriate storage condition.

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INCREMENT SERVICES

Magazine Citations
Nature Medicine Angiotensin-Converting Enzyme 2 Overexpression Remarkably Ameliorated Glomerular Injury in a Rat Model of Diabetic Nephropathy: A Comparison with ACE Inhibition MolMed: 10_11_liu
Journal of Cancer Therapy Plasma Levels of Angiotensin-Converting Enzymes 1 and 2 and AGTR2 (T1247G and A5235G) Gene Polymorphisms Are Associated to Breast Cancer Progression. Scirp: Source
Reprod Toxicol.? ACE2 activation by xanthenone prevents leptin-induced increases in blood pressure and proteinuria during pregnancy in Sprague-Dawley rats Pubmed:25205467
Intensive Care Medicine Experimental Angiotensin converting enzymes in patients with acute respiratory distress syndrome Content: 3
Journal of Chromatography A Application of a new procedure for liquid chromatography/mass spectrometry profiling of plasma amino acid-related metabolites and untargeted shotgun proteomics to identify mechanisms and biomarkers of calcific aortic stenosis S0021967317311858
Journal of Clinical Pharmacy and Therapeutics Relationship between genetic variants of ACE2 gene and circulating levels of ACE2 and its metabolites. pubmed:28895159
Innate Immunity  Lipoxin A attenuates LPS-induced acute lung injury via activation of the ACE2-Ang-(1-7)-Mas axis Pubmed:29969931
Journal of Surgical Research Impact of angiotensin-converting enzyme 2 levels on postoperative pneumonia after esophagectomy Pubmed:29506841
canadian journal of physiology and pharmacology Could cardioprotective effect of ACE2 activator “diminazene aceturate” is more potent than ACE inhibitor “Enalapril” on acute myocardial infarction in rats? Pubmed: 30840489
Intensive Care Medicine Angiotensin-converting enzymes in acute respiratory distress syndrome
BioMed Research International AVE 0991 Attenuates Pyroptosis and Liver Damage after Heatstroke by Inhibiting the ROS-NLRP3 Inflammatory Signalling Pathway
Heart and vessels Relationship between circulating levels of angiotensin-converting enzyme 2-angiotensin-(1–7)-MAS axis and coronary heart disease Pubmed: 31359146
Medicine Alteration and association between serum ACE2/angiotensin (1-7)/Mas axis and oxidative stress in chronic kidney disease: A pilot study Pubmed: 32756181
American Journal of Respiratory and Critical Care Medicine Equilibrium Angiotensin Metabolite Profiling in Patients with Acute Respiratory Distress Syndrome Indicates Angiotensin Converting Enzyme Inhibition Pubmed: 32628511
Clinical Research in Cardiology Determinants of soluble angiotensin-converting enzyme 2 concentrations in adult patients with complex congenital heart disease 33280062
Clinical & Translational Immunology Pattern of circulating SARS©\CoV©\2©\specific antibody©\secreting and memory B©\cell generation in patients with acute COVID©\19 33552508
researchsquare Novel transgenic mice with Cre-dependent co-expression of GFP and human ACE2: a safe tool for study of COVID-19 pathogenesis 33855640
Viruses Different Neutralization Sensitivity of SARS-CoV-2 Cell-to-Cell and Cell-Free Modes of Infection to Convalescent Sera
Sci Rep Immunoreactivity of the SARS-CoV-2 entry proteins ACE-2 and TMPRSS-2 in murine models of hormonal manipulation, ageing, and cardiac injury 34907257
European Journal of Clinical Investigation Angiotensin‐converting enzyme 2 and Transmembrane protease serine 2 in female and male patients with end‐stage kidney disease Pubmed:35366343
Metabolites Ramipril Reduces Acylcarnitines and Distinctly Increases Angiotensin-Converting Enzyme 2 Expression in Lungs of Rats Pubmed:35448480
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