L-Aspartic acid β-7-amido-4-methylcoumarin
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L-Aspartic acid β-7-amido-4-methylcoumarin

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Category
L-Amino Acids
Catalog number
BAT-004114
CAS number
133628-73-6
Molecular Formula
C14H14N2O5
Molecular Weight
290.23
L-Aspartic acid β-7-amido-4-methylcoumarin
IUPAC Name
(2S)-2-amino-4-[(4-methyl-2-oxochromen-7-yl)amino]-4-oxobutanoic acid
Synonyms
L-Asp(AMC)-OH; L-Aspartic acid β-(7-amido-4-methylcoumarin)
Appearance
White crystalline powder
Purity
≥ 99% (HPLC)
Density
1.452±0.06 g/cm3
Melting Point
193-198 °C
Boiling Point
610.0±55.0 °C
Storage
Store at 2-8 °C
InChI
InChI=1S/C14H14N2O5/c1-7-4-13(18)21-11-5-8(2-3-9(7)11)16-12(17)6-10(15)14(19)20/h2-5,10H,6,15H2,1H3,(H,16,17)(H,19,20)/t10-/m0/s1
InChI Key
ARZPQBJTLVVDNP-JTQLQIEISA-N
Canonical SMILES
CC1=CC(=O)OC2=C1C=CC(=C2)NC(=O)CC(C(=O)[O-])[NH3+]

L-Aspartic acid β-7-amido-4-methylcoumarin is a synthetic compound that belongs to the class of aspartic acid derivatives. Structurally, it is characterized by the presence of an amido group and a coumarin moiety linked to the beta position of aspartic acid. Coumarin derivatives are widely studied for their diverse biological activities and applications, and when introduced with aspartic acid, these compounds can exhibit unique biochemical properties. The presence of coumarin usually imparts fluorescent properties which make it useful in various diagnostic and analytical techniques. This compound is primarily recognized for its role in scientific and medical research as a molecular probe due to these intrinsic properties.

One of the key applications of L-Aspartic acid β-7-amido-4-methylcoumarin is in fluorescence microscopy and imaging. The coumarin moiety in the compound fluoresces under specific wavelengths of light, making it an excellent candidate for use in tracking biological processes in live cells. These fluorescent properties are especially valuable for imaging studies which require real-time analysis of cellular behavior and interactions. Researchers utilize this compound to tag proteins or other biomolecules, allowing them to observe the dynamic processes in cellular mechanisms, including protein synthesis, localization, and degradation pathways.

Another important application is its use in enzyme activity assays, particularly for enzymes that cleave peptide bonds. L-Aspartic acid β-7-amido-4-methylcoumarin serves as a substrate that releases a fluorescent signal upon enzymatic action. This makes it particularly useful for studying enzymes such as peptidases or proteases, where its cleavage results can be quantified by fluorescence. This type of application is crucial in drug discovery and diagnostics, where understanding enzyme mechanisms can lead to the development of therapeutic inhibitors or activators. By providing a means to quantitatively assess enzyme activity, this compound enhances the identification of enzyme function and regulation.

L-Aspartic acid β-7-amido-4-methylcoumarin is also employed in drug discovery and pharmacological research. Its fluorescent properties enable researchers to design assays that test the efficacy and binding characteristics of drug molecules. By using the compound as a marker, it is possible to visualize drug interactions at the molecular level, evaluating how potential pharmaceuticals interact with specific targets. This approach is key for developing new drugs, as it allows for the high-throughput screening of compounds and the study of potential side effects or off-target interactions, leading to more effective and safer medications.

Lastly, the compound finds its use in environmental sciences, particularly in monitoring and detecting pollutants. The sensitivity of its fluorescent response allows for the detection of trace amounts of toxic substances in soil and water. By binding to specific pollutants, L-Aspartic acid β-7-amido-4-methylcoumarin offers a visual indicator of contamination presence and concentration levels. This application is vital for environmental monitoring and regulation, enabling researchers and authorities to carry out quantitative assessments of pollution and its sources, ultimately guiding cleanup and policy efforts. The compound’s versatility and sensitivity make it an invaluable tool in preserving environmental and public health.

1. A fluorometric assay for glycosylasparaginase activity and detection of aspartylglycosaminuria
I T Mononen, V M Kaartinen, J C Williams Anal Biochem. 1993 Feb 1;208(2):372-4. doi: 10.1006/abio.1993.1063.
Recent experimental work on the mechanism of action of glycosylasparaginase suggests that the enzyme specifically reacts toward the L-asparagine or L-aspartic acid moiety of its substrates. Based on this, a new sensitive assay for glycosylasparaginase activity has been developed using L-aspartic acid beta-(7-amido-4-methylcoumarin) as substrate. Release of 7-amino-4-methylcoumarin was determined fluorometrically. At pH 7.5, Km = 93 microM, and as little as 1 ng of glycosylasparaginase could be detected with the assay. Hydrolysis of the substrate was inhibited by diazo-oxonorvaline, a specific inhibitor of glycosylasparaginase. In biological samples, the fluorometric assay is 40-100 times more sensitive than other published methods for glycosylasparaginase. This new assay enables a rapid enzymatic diagnosis of aspartylglycosaminuria--a genetic deficiency of glycosylasparaginase activity--with leukocyte and fibroblast samples.
2. A fluorometric assay for L-asparaginase activity and monitoring of L-asparaginase therapy
P Ylikangas, I Mononen Anal Biochem. 2000 Apr 10;280(1):42-5. doi: 10.1006/abio.2000.4500.
The antineoplastic enzyme L-asparaginase is commonly used for the induction of remission in acute lymphoblastic leukemia (ALL). There is no simple method available for measuring the activity of this highly toxic drug. We incubated L-asparaginase from Erwinia chrysanthemi with L-aspartic acid beta-(7-amido-4-methylcoumarin) and measured the release of 7-amino-4-methylcoumarin fluorometrically for 30-300 min. The rate of the hydrolysis of the substrate was linear over a 50-fold range of the concentration of the enzyme. With increasing substrate concentration, the enzyme showed a saturable kinetic pattern with V(max) of 0.547 (SD 0.059) microM/min/mg of enzyme (n = 3) and Km of 0.302 (SD 0.095) mM (n = 3). This assay enables rapid analysis of L-asparaginase activity in biological samples and it can be used, for example, for monitoring of L-asparaginase activity in serum of ALL patients during their L-asparaginase therapy.
3. Enzymatic diagnosis of aspartylglycosaminuria by fluorometric assay of glycosylasparaginase in serum, plasma, or lymphocytes
I Mononen, T Mononen, P Ylikangas, V Kaartinen, K Savolainen Clin Chem. 1994 Mar;40(3):385-8.
Serum, plasma, and lymphocytes from aspartylglycosaminuria (AGU) patients and carriers and from normal controls were incubated with a fluorescent glycosylasparaginase substrate, L-aspartic acid beta-(7-amido-4-methylcoumarin), and the release of 7-amino-4-methylcoumarin was measured fluorometrically after incubation for 1-4 h. The mean glycosylasparaginase (EC 3.5.1.26) activity in normal serum, plasma, and lymphocytes was 20.2 (SD 5.0) mU/L (n = 24), 17.5 (SD 5.0) mU/L (n = 24), and 242 (SD 108) mU/g protein (n = 17), respectively. The corresponding values in the Finnish AGU patients were 0.7 (SD 0.4) mU/L (n = 10), 0.3 (SD 0.3) mU/L (n = 10), and 6.0 (SD 4.6) mU/g protein (n = 7). No overlapping values were obtained between the AGU patients and the carriers in any of the samples, but the values between the carriers and controls were overlapping in 28 of 29 serum, 22 of 29 plasma, and 4 of 21 lymphocyte samples. Thus, the fluorometric glycosylasparaginase assay in various blood samples allows specific detection of the enzyme defect in AGU, but cannot be used for reliable detection of carriers of the disease.
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