Angiotensin I Converting Enzyme 2, (ACE-2) Substrate

Product Name: Angiotensin I Converting Enzyme 2, (ACE-2) Substrate

Sequence One Letter Code: Mca-APK(Dnp)

Sequence Three Letter Code: Mca-Ala-Pro-Lys(Dnp)-OH

Cas No: 305336-82-7

Chemical Formula:C30H34N6O10

Molecular Weight: 696.7

Purity: 95%

Form: Lyophilized

Storage Conditions: - 20 °C Protected from light

Research Area: peptide substrate

SMILES: C[C@@H](C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCCCNC2=C(C=C(C=C2)[N+](=O)[O-])[N+](=O)[O-])C(=O)O)NC(=O)CC3=CC(=O)OC4=C3C=CC(=C4)OC

IUPAC: (2S)-6-(2,4-dinitroanilino)-2-[[(2S)-1-[(2S)-2-[[2-(7-methoxy-2-oxochromen-4-yl)acetyl]amino]propanoyl]pyrrolidine-2-carbonyl]amino]hexanoic acid

INCHIKEY: INOZJECNSBDPGT-WDNCENIBSA-N

INCHI:

InChI=1S/C32H36N6O12/c1-18(34-28(39)14-19-15-29(40)50-27-17-21(49-2)9-10-22(19)27)31(42)36-13-5-7-25(36)30(41)35-24(32(43)44)6-3-4-12-33-23-11-8-20(37(45)46)16-26(23)38(47)48/h8-11,15-18,24-25,33H,3-7,12-14H2,1-2H3,(H,34,39)(H,35,41)(H,43,44)/t18-,24-,25-/m0/s1

Conjugation: Conjugated

Conjugation Type: Double dyes

Code Nacres: NA.26

Application: Angiotensin I–Converting Enzyme 2 (ACE-2) Substrate is a fluorogenic peptide engineered for quantitative assessment of ACE-2 catalytic activity in vitro. Upon enzymatic cleavage, the quenched fluorophore is released, producing a robust fluorescence signal detectable at Ex/Em 325/393 nm. Complete hydrolysis of 0.04 mM substrate yields approximately a 300-fold signal increase over background, enabling sensitive kinetic analysis. ACE-2 is a zinc-dependent carboxypeptidase that counterbalances the classical renin–angiotensin system by converting angiotensin II into angiotensin-(1–7). This substrate supports enzyme characterization, determination of kinetic parameters, and screening of ACE-2 modulators. It is widely applied in cardiovascular research, enzymology studies, and investigations of renin–angiotensin system regulation under physiological and pathological conditions.

Current Research: Angiotensin I–Converting Enzyme 2 (ACE-2) Substrate is a fluorogenic peptide designed for sensitive and quantitative measurement of ACE-2 catalytic activity in vitro. The substrate incorporates a quenched fluorophore that becomes highly fluorescent upon enzymatic cleavage, generating a signal detectable at Ex/Em 325/393 nm. Complete hydrolysis of 0.04 mM substrate produces approximately a 300-fold increase in fluorescence over background, enabling robust kinetic analysis with high signal-to-noise performance. ACE-2 in the Renin–Angiotensin System ACE-2 is a zinc-dependent monocarboxypeptidase that plays a central regulatory role in the renin–angiotensin system (RAS). While the classical RAS axis involves conversion of angiotensin I to angiotensin II via ACE, leading to vasoconstriction and pro-inflammatory signaling, ACE-2 counterbalances this pathway by converting angiotensin II into angiotensin-(1–7). The latter peptide exerts vasodilatory, anti-inflammatory, and anti-fibrotic effects through activation of the Mas receptor. By modulating angiotensin peptide balance, ACE-2 influences: Blood pressure regulation Vascular tone Cardiac remodeling Renal function Inflammatory responses Given its protective role, quantitative assessment of ACE-2 activity is critical in cardiovascular and metabolic research. Mechanism of Fluorogenic Detection The ACE-2 Substrate utilizes a fluorescence resonance energy transfer (FRET)–based or quenched fluorophore system. In its intact form, fluorescence emission is suppressed due to proximity-dependent quenching. Upon ACE-2–mediated cleavage at the designated carboxypeptidase recognition site, the fluorophore is released from quenching constraints, producing a measurable increase in fluorescence intensity. This assay configuration offers several analytical advantages: Continuous, real-time monitoring of enzyme activity High sensitivity due to strong fluorescence amplification Direct correlation between cleavage rate and fluorescence intensity Homogeneous assay format without separation steps The spectral profile (325/393 nm) is compatible with standard fluorescence spectrophotometers and microplate readers. Applications in Enzyme Characterization The substrate supports detailed enzymatic profiling of recombinant or purified ACE-2, including: Determination of kinetic parameters (K_m, V_max, catalytic efficiency) Comparative analysis of ACE-2 variants or mutants Evaluation of enzyme stability and activity under varying conditions Time-dependent cleavage assays Because ACE-2 is a zinc metalloprotease, the substrate is also useful for studying metal ion dependency and catalytic mechanism. Screening of ACE-2 Modulators Pharmacological modulation of ACE-2 is of interest in cardiovascular disease, pulmonary disorders, and inflammatory pathologies. The fluorogenic substrate enables: High-throughput screening of small-molecule activators or inhibitors IC₅₀ determination for candidate modulators Structure–activity relationship (SAR) studies Assessment of selective targeting relative to ACE The strong fluorescence amplification facilitates detection of subtle changes in catalytic activity. Cardiovascular and Pathophysiological Research ACE-2 plays a protective role in conditions such as hypertension, heart failure, and vascular injury. Quantitative measurement of ACE-2 activity is essential for understanding how shifts in angiotensin peptide balance contribute to disease progression. The substrate is widely used in: Cardiovascular enzyme assays Studies of RAS pathway regulation Renal and pulmonary physiology research Models of inflammatory and fibrotic signaling Experimental Advantages High fluorescence amplification (~300-fold signal increase) Sensitive detection at low substrate concentrations Compatible with microplate-based high-throughput workflows Suitable for purified enzyme and biological sample analysis Enables reproducible kinetic evaluation Research Significance Angiotensin I–Converting Enzyme 2 (ACE-2) Substrate provides a reliable and sensitive platform for investigating ACE-2 enzymatic function and regulatory mechanisms within the renin–angiotensin system. By enabling precise quantification of catalytic activity, it supports mechanistic studies of cardiovascular regulation, enzymology, and pharmacological modulation in both physiological and disease-relevant experimental models.