Angiotensin II Converting Enzyme (ACE2) Inhibitor Screening Kit (BN00569)
- SKU:
- BN00569
Description
Angiotensin II Converting Enzyme (ACE2) Inhibitor Screening Kit
Angiotensin II converting enzyme (ACE2, EC 3.4.17.23), a carboxypeptidase, is part of the renin-angiotensin system (RAS) that controls regulation of blood pressure by cleaving the C-terminal dipeptide of Angiotensin II to convert it into Angiotensin 1-7. It also cleaves Angiotensin I to produce Ang 1-9, of unknown function. ACE2 is a receptor of human coronaviruses, such as SARS and HCoV-NL63. It is expressed on the vascular endothelial cells of heart and kidney. The inhibitors of ACE2 could be able to regulate hypertension by changing vascular permeability. Screening for small molecule and peptide inhibitors might also help in finding treatment for coronavirus mediated infection. Assay Genie's ACE2 Inhibitor Screening Kit can be used to screen for potent inhibitors of ACE2 activity, it utilizes the ability of an active ACE2 to cleave a synthetic MCA based peptide substrate to release a free fluorophore. The released MCA can be easily quantified using a fluorescence microplate reader. In the presence of an ACE2 specific inhibitor, the enzyme loses its peptidase activity which results in decrease of fluorescence intensity. This assay kit is simple and can be used to identify and characterize ACE2 inhibitors in a high-throughput format.
Figure: Inhibition of ACE2 activity by ACE2 Inhibitor, IC 50 = 33.0 ± 0.6 nM (n = 3). Assay was performed following the kit protocol.
| Key Information | Description |
| Product SKU | BN00569 |
| Size | 100 assays |
| Detection Method | Fluorescence (Ex/Em = 320/420 nm) |
| Applications | Screening/characterizing/studying ACE2 inhibitors/activators |
| Features and Benefits |
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| Kit Components |
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| Storage Conditions | -20°C |
| Shipping Conditions | Gel Pack |
| USAGE | For Research Use Only! Not For Use in Humans. |
ACE2 function
Carboxypeptidase which converts angiotensin I to angiotensin 1-9, a peptide of unknown function, and angiotensin II to angiotensin 1-7, a vasodilator (PubMed:10969042, PubMed:10924499, PubMed:11815627). Also able to hydrolyze apelin-13 and dynorphin-13 with high efficiency (PubMed:11815627). By cleavage of angiotensin II, may be an important regulator of heart function (PubMed:10969042, PubMed:10924499). By cleavage of angiotensin II, may also have a protective role in acute lung injury (By similarity). Plays an important role in amino acid transport by acting as binding partner of amino acid transporter SL6A19 in intestine, regulating trafficking, expression on the cell surface, and its catalytic activity (PubMed:18424768, PubMed:19185582).
ACE2 & the coronavirus
Angiotensin-converting enzyme 2 (ACE2) has been determined to be a functional receptor which facilitates the uptake of SARS-CoV-2 into host cells. (Source) ACE2 is expressed in multiple locations and cell types, notably including alveolar epithelial cells, surface enterocytes within the small intestine, and heart and kidney endothelial cells.
Coronavirus entry into susceptible cells is mediated by the S1 subunit of the coronavirus transmembrane ‘spike’ glycoprotein(commonly abbreviated ‘S’). (Source) S1 contains a receptor binding domain at residues 318–510, which recognises and binds to the LYS341 residue of ACE2 with high affinity. (Source) The affinity of CoV-2 is higher than that of CoV. (Source)
Increased binding affinity to human ACE2 has been correlated with an increase in transmission efficiency in previous outbreaks of SARS-CoV. (Source) Antibodies produced against SARS-CoV-2 target the S2 subunit, which is responsible for the fusion of viral and host membranes. Thus, neutralising the S2 subunit inhibits viral entry into vulnerable cells. (Source) Polyclonal antibodies against the S protein of SARS-CoV have been found to significantly attenuate the ability of SARS-CoV-2 to enter cells via human ACE2. (Source)
| ACE2 | Key information |
| Uniprot | |
| Gene name | ACE2 |
| Alternative names | Angiotensin-converting enzyme 2, ACE-related carboxypeptidase, Angiotensin-converting enzyme homolog, ACEH, Metalloprotease MPROT15 |
| Subcellular location | Extracellular region or secreted, cell membrane, cytoplasm |
| Tissue Specificity | Expressed in endothelial cells from small and large arteries, and in arterial smooth muscle cells (at protein level) (PubMed:15141377). Expressed in lung alveolar epithelial cells, enterocytes of the small intestine, Leydig cells and Sertoli cells (at protein level) (PubMed:15141377). Expressed in the renal proximal tubule and the small intestine (at protein level) (PubMed:18424768). Expressed in heart, kidney, testis, and gastrointestinal system (PubMed:10969042, PubMed:10924499, PubMed:15231706, PubMed:12459472, PubMed:15671045). |
| Induction | Up-regulated in failing heart. |
ACE 2 Activity Assay Kit - Protocol
| Step | Procedure |
| ACE2 Enzyme Working Solution Preparation: | Add 198 µl of ACE2 Dilution Buffer to the ACE2 enzyme vial. The diluted enzyme can EC, S, SC and IC
Background Control (BC)
Mix well and add 50 μl/well into desired wells in a 96-well microtiter plate |
| Screening Compounds, Inhibitor Control & Enzyme Control Preparations: | Dissolve candidate inhibitors at 100X highest final test concentration using preferred solvent. Dilute to 10X the desired test concentration with ACE2 Assay Buffer. Add 10 μl test inhibitors (Sample, S) or ACE2 Assay Buffer (EC and BC). Prepare Inhibitor Control by adding 50 µl ACE2 Assay Buffer to the vial containing ACE2 Inhibitor, mix. For Inhibitor Control (IC), add 10 μl ACE2 Inhibitor into ACE2 enzyme containing well(s). Incubate at Room temperature (RT) for 15 min. |
| ACE2 Substrate Mix: | Prepare enough reagents for the number of assays to be performed. For each well, prepare 40 μl of the Substrate Mix:
Mix & add 40 µl of ACE2 Substrate Mix into Background Control, Enzyme Control/Solvent Control, Inhibitor Control & Sample (S) wells. Mix well. |
| Measurement: | Measure fluorescence (Ex/Em = 320/420 nm) in a kinetic mode for 1 hr at room temperature. |
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