ACE2 Protein, Rat (HEK293, His)

ACE2 is an important carboxypeptidase in the renin-angiotensin system that complexly regulates cardiovascular homeostasis. It specifically converts angiotensin I to angiotensin 1-9 and angiotensin II to angiotensin 1-7, exerting antihypertrophic and vasodilatory effects. ACE2 Protein, Rat (HEK293, His) is the recombinant rat-derived ACE2 protein, expressed by HEK293 , with C-His labeled tag.

ACE2, an essential counter-regulatory carboxypeptidase within the renin-angiotensin hormone system, stands as a pivotal regulator orchestrating blood volume, systemic vascular resistance, and overall cardiovascular homeostasis. This multifunctional enzyme displays remarkable specificity in its actions, converting angiotensin I to angiotensin 1-9 and angiotensin II to angiotensin 1-7. These biologically active peptides exhibit anti-hypertrophic effects in cardiomyocytes and act as beneficial vasodilators, counteracting the vasoconstrictor properties of angiotensin II. Beyond its role in the angiotensin system, ACE2 demonstrates versatility by cleaving various vasoactive peptides, including neurotensin, kinetensin, and des-Arg bradykinin. Notably, ACE2 is intricately involved in amino acid transport, serving as a binding partner for the amino acid transporter SLC6A19 in the intestine. Through its involvement in trafficking, surface expression, and catalytic activity modulation, ACE2 plays a crucial role in the regulation of amino acid transport processes.

Measured by its ability to cleave a fluorogenic peptide substrate, Mca-YVADAPK(Dnp)-OH. The specific activity is 20028.43 pmol/min/µg, as measured under the described conditions.

Materials
Detection buffer: 50 mM Tris, 1M Nacl, pH 7.0
ACE2 Protein, Rat (HEK293, His) (HY-P72809)
Substrate: Mca-YVADAPK(Dnp) -OH was solubilized to 2 mM with DMSO
Standard: MCA-Pro-Leu-OH

Procedure
1. Dilute MCA-Pro-Leu-OH to 50, 25, 12.5, 6.25, 3.125, 1.5625, 0.78, 0.39, 0.2, 0 μmol/L with assay buffer, and then add 100 μL of each buffer into wells to start the reaction. The excitation and emission wavelengths were 320 nm and 405 nm , respectively, and read in kinetic mode for 5 min. The standard curve was made with the concentration of the standard as the horizontal coordinate and the measured RFU as the vertical coordinate, and the standard curve formula was obtained.
2. Dilute Rat ACE2 to 0.05, 0.5 , 5 μg/mL with assay buffer.
3. The substrate was diluted to 20 μM with assay buffer.
4. Add different concentrations of ACE2 to the black well plate at 50 μL per well and start the reaction by adding 50 μL of 20 μM substrate. Blank wells were added with 50 μL of assay buffer and 50 μL of 20 μM substrate.
5. Read in kinetic mode for 5 minutes at 320 nm and 405 nm excitation and emission wavelengths, respectively.
6. Calculate the specific activity:

Rat

HEK293

C-His

Q5EGZ1 (Q18-T740)

302668  [NCBI]

Extracellular Domain

QSLIEEKAESFLNKFNQEAEDLSYQSSLASWNYNTNITEENAQKMNEAAAKWSAFYEEQSKIAQNFSLQEIQNATIKRQLKALQQSGSSALSPDKNKQLNTILNTMSTIYSTGKVCNSMNPQECFLLEPGLDEIMATSTDYNRRLWAWEGWRAEVGKQLRPLYEEYVVLKNEMARANNYEDYGDYWRGDYEAEGVEGYNYNRNQLIEDVENTFKEIKPLYEQLHAYVRTKLMEVYPSYISPTGCLPAHLLGDMWGRFWTNLYPLTTPFLQKPNIDVTDAMVNQSWDAERIFKEAEKFFVSVGLPQMTPGFWTNSMLTEPGDDRKVVCHPTAWDLGHGDFRIKMCTKVTMDNFLTAHHEMGHIQYDMAYAKQPFLLRNGANEGFHEAVGEIMSLSAATPKHLKSIGLLPSNFQEDNETEINFLLKQALTIVGTLPFTYMLEKWRWMVFQDKIPREQWTKKWWEMKREIVGVVEPLPHDETYCDPASLFHVSNDYSFIRYYTRTIYQFQFQEALCQAAKHDGPLHKCDISNSTEAGQKLLNMLSLGNSGPWTLALENVVGSRNMDVKPLLNYFQPLFVWLKEQNRNSTVGWSTDWSPYADQSIKVRISLKSALGKNAYEWTDNEMYLFRSSVAYAMREYFSREKNQTVPFGEADVWVSDLKPRVSFNFFVTSPKNVSDIIPRSEVEEAIRMSRGRINDIFGLNDNSLEFLGIYPTLKPPYEPPVT

Approximately 105-115 kDa, based on SDS-PAGE under reducing conditions.

• 
  ≥ 95%, as determined by reducing SDS-PAGE.

Lyophilized powder.

Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.4.

<1 EU/μg, determined by LAL method.

It is not recommended to reconstitute to a concentration less than 100 μg/mL in ddH₂O. For long term storage it is recommended to add a carrier protein (0.1% BSA, 5% HSA, 10% FBS or 5% Trehalose).

Stored at -20°C for 2 years from date of receipt. After reconstitution, it is stable at 4°C for 1 week or -20°C for longer (with carrier protein). It is recommended to freeze aliquots at -20°C or -80°C for extended storage.

Room temperature in continental US; may vary elsewhere.