Angiotensin I Converting Enzyme 2, ACE-2/Caspase-1 Substrate

Product Name: Angiotensin I Converting Enzyme 2, ACE-2/Caspase-1 Substrate

Sequence One Letter Code: Mca-YVADAPK(Dnp)

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

Cas No: 189696-01-3

Chemical Formula:C53H64N10O19

Molecular Weight: 1145.2

Purity: 95%

Form: Lyophilized

Storage Conditions: - 20 °C Protected from light

Research Area: Inflammation and Immunology Research

SMILES: C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](C)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)[C@H](C(C)C)NC(=O)[C@H](CC3=CC=C(C=C3)O)NC(=O)CC4=CC(=O)OC5=C4C=CC(=C5)OC

IUPAC: 2S)-2-[[(2S)-1-[(2S)-2-[[(2S)-3-carboxy-2-[[(2S)-2-[[(2S)-2-[[(2S)-3-(4-hydroxyphenyl)-2-[[2-(7-methoxy-2-oxochromen-4-yl)acetyl]amino]propanoyl]amino]-3-methylbutanoyl]amino]propanoyl]amino]propanoyl]amino]propanoyl]pyrrolidine-2-carbonyl]amino]-6-(2,4-dinitroanilino)hexanoic acid

INCHIKEY: XLXKHBNLWUCNOC-XUFBPUFYSA-N

INCHI:

InChI=1S/C53H64N10O19/c1-27(2)46(60-49(71)38(21-30-11-14-33(64)15-12-30)57-43(65)22-31-23-45(68)82-42-25-34(81-5)16-17-35(31)42)51(73)55-28(3)47(69)59-39(26-44(66)67)48(70)56-29(4)52(74)61-20-8-10-40(61)50(72)58-37(53(75)76)9-6-7-19-54-36-18-13-32(62(77)78)24-41(36)63(79)80/h11-18,23-25,27-29,37-40,46,54,64H,6-10,19-22,26H2,1-5H3,(H,55,73)(H,56,70)(H,57,65)(H,58,72)(H,59,69)(H,60,71)(H,66,67)(H,75,76)/t28-,29-,37-,38-,39-,40-,46-/m0/s1

Conjugation: Conjugated

Conjugation Type: Double dyes

Code Nacres: NA.26

Application: ACE-2 / Caspase-1 Substrate is a fluorogenic peptide designed for sensitive measurement of caspase-1 proteolytic activity. Cleavage of the Mca-labeled FRET substrate releases fluorescence detectable at Ex/Em 325/393 nm, enabling quantitative kinetic analysis. Caspase-1 is a cysteine protease central to inflammasome activation and the maturation of pro-inflammatory cytokines such as IL-1β and IL-18. This substrate supports biochemical characterization of caspase-1 activity and evaluation of inflammasome modulators in vitro. Its defined cleavage sequence and reliable fluorescence output make it suitable for inhibitor screening, pathway dissection, and mechanistic studies of inflammatory signaling. The substrate is broadly applied in immunology, inflammation biology, and innate immune response research models.

Current Research: ACE-2 / Caspase-1 Substrate is a fluorogenic peptide engineered for sensitive and quantitative detection of caspase-1 proteolytic activity in biochemical and cell-based assays. The peptide incorporates an Mca (7-methoxycoumarin-4-acetyl) fluorophore within a FRET-based design. In the intact substrate, fluorescence is quenched; upon specific cleavage by caspase-1, separation of the fluorophore from the quenching moiety generates a measurable fluorescence signal at Ex/Em 325/393 nm. This configuration enables continuous kinetic monitoring of enzyme activity with high sensitivity and low background interference. Caspase-1 and Inflammasome Biology Caspase-1 is a cysteine protease activated within multiprotein inflammasome complexes, including NLRP3, AIM2, and NLRC4 inflammasomes. Upon activation, caspase-1 cleaves pro–IL-1β and pro–IL-18 into their mature, biologically active forms, driving inflammatory signaling cascades. In addition to cytokine processing, caspase-1 mediates pyroptosis through cleavage of gasdermin D, resulting in membrane pore formation and inflammatory cell death. Because of its central role in innate immune activation, caspase-1 is implicated in numerous inflammatory and autoimmune conditions, including sepsis, gout, metabolic syndrome, neuroinflammation, and chronic inflammatory disorders. Precise quantification of caspase-1 activity is therefore essential for understanding inflammasome dynamics and evaluating potential therapeutic modulators. Mechanism of Fluorogenic Detection The ACE-2 / Caspase-1 Substrate contains a defined peptide sequence recognized and cleaved by active caspase-1. In its uncleaved state, fluorescence emission from the Mca fluorophore is suppressed due to proximity-dependent quenching. Proteolytic cleavage disrupts this interaction, resulting in an increase in fluorescence intensity proportional to enzymatic activity. This FRET-based approach offers several advantages: Real-time monitoring of caspase-1 activity Quantitative kinetic analysis (V_max, K_m determination) Minimal assay interference Homogeneous assay format without separation steps The excitation/emission profile (325/393 nm) is compatible with standard fluorescence microplate readers and spectrofluorometers, facilitating integration into routine enzymology workflows. Applications in Inflammasome Research This substrate is widely applied in: Biochemical characterization of recombinant caspase-1 Measurement of inflammasome activation in cell lysates Evaluation of NLRP3 pathway activation Screening of small-molecule caspase-1 inhibitors Comparative analysis of inflammasome modulators In cell-based models, the substrate enables quantitative assessment of caspase-1 activation following stimulation with canonical inflammasome activators such as ATP, nigericin, or pathogen-associated molecular patterns (PAMPs). This supports mechanistic dissection of upstream signaling events and downstream cytokine maturation pathways. Inhibitor Screening and Drug Development Given the therapeutic interest in targeting inflammasome signaling, caspase-1 inhibitors are under investigation for inflammatory and metabolic diseases. The fluorogenic substrate provides a reliable platform for: High-throughput inhibitor screening Determination of IC₅₀ values Structure–activity relationship (SAR) studies Selectivity profiling against related caspases Its defined cleavage sequence ensures specificity for caspase-1–dependent proteolysis, enabling accurate pharmacological evaluation. Experimental Advantages Sensitive fluorescence detection Continuous kinetic readout Defined caspase-1 cleavage sequence Suitable for biochemical and cell-based assays Compatible with standard fluorescence instrumentation The substrate’s robust fluorescence response and reproducibility make it a dependable tool for investigating inflammatory signaling pathways. Research Impact By enabling precise quantification of caspase-1 activity, ACE-2 / Caspase-1 Substrate supports detailed analysis of inflammasome biology, cytokine maturation, and pyroptotic signaling. Its utility in enzymatic profiling and inhibitor screening makes it particularly valuable in immunology, inflammation research, and translational studies focused on innate immune modulation.