L-Alanine 4-nitroanilide hydrochloride
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L-Alanine 4-nitroanilide hydrochloride

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Category
L-Amino Acids
Catalog number
BAT-003947
CAS number
31796-55-1
Molecular Formula
C9H11N3O3·HCl
Molecular Weight
245.70
L-Alanine 4-nitroanilide hydrochloride
IUPAC Name
(2S)-2-amino-N-(4-nitrophenyl)propanamide;hydrochloride
Synonyms
L-Ala-pNA HCl; (S)-2-Amino-N-(4-nitrophenyl)propanamide hydrochloride; (2S)-2-amino-N-(4-nitrophenyl)propanamide hydrochloride
Appearance
Light yellowish powder
Purity
≥ 99% (HPLC)
Melting Point
181-186 °C
Boiling Point
443.4 °C at 760 mmHg
Storage
Store at 2-8 °C
InChI
InChI=1S/C9H11N3O3.ClH/c1-6(10)9(13)11-7-2-4-8(5-3-7)12(14)15;/h2-6H,10H2,1H3,(H,11,13);1H/t6-;/m0./s1
InChI Key
YEXRLSXNWLNHQR-RGMNGODLSA-N
Canonical SMILES
CC(C(=O)NC1=CC=C(C=C1)[N+](=O)[O-])N.Cl

L-Alanine 4-nitroanilide hydrochloride is a synthetic substrate used primarily in biochemical research. Here are some key applications of L-Alanine 4-nitroanilide hydrochloride:

Enzyme Kinetics: L-Alanine 4-nitroanilide hydrochloride is used as a chromogenic substrate in enzyme assays to study peptidase activity. When cleaved by specific enzymes, it releases a chromophore that can be quantitatively measured. This helps in understanding enzyme kinetics, substrate specificity, and inhibitor screening.

Protein Profiling: In proteomics, L-Alanine 4-nitroanilide hydrochloride facilitates the identification of peptidases in complex biological samples. By monitoring the substrate's cleavage, researchers can profile the presence and activity levels of these enzymes. This approach is instrumental in understanding protein function and disease pathology.

Pharmaceutical Development: L-Alanine 4-nitroanilide hydrochloride is also valuable in drug discovery, where it is used to screen potential inhibitors for peptidases. Identifying effective inhibitors can lead to the development of new therapeutic agents. The substrate's sensitivity and specificity make it a useful tool in high-throughput screening.

Diagnostic Assays: This compound is utilized in clinical diagnostics to measure enzyme activity in biological fluids like blood or urine. By assessing the rate of substrate cleavage, clinicians can diagnose and monitor conditions associated with abnormal enzyme activity. This provides a non-invasive method for disease detection and management.

1. Visualization of protein digestion in the midgut of the acarid mite Lepidoglyphus destructor
Tomas Erban, Jan Hubert Arch Insect Biochem Physiol. 2011 Oct;78(2):74-86. doi: 10.1002/arch.20441. Epub 2011 Aug 4.
The ingestion of chromogenic or fluorescent substrates for protease detection enables the visualization of digestive processes in mites in vivo due to their transparent bodies. The substrates for protease detection were offered to Lepidoglyphus destructor, and the resulting signals were observed in specimens under a compound microscope. The protease activity was successfully localized using chromogenic substrates (azoalbumin, AAPpNA, SAAPFpNA, elastin-orcein, SA(3) pNA, ZRRpNA, ArgpNA, and MAAPMpNA) and fluorescent substrates (casein-fluorescein, albumin-fluorescein, AAPAMC, BAAMC, ZRRAMC, ArgAMC, and AGPPPAMC). No activity was detected using the keratin azure and BApNA substrates. In the mesodeum, trypsin-like activity generated by hydrolysis of the BApNA substrate was not observed, but the BAAMC substrate allowed the visualization of trypsin-like activity in food boli in the posterior mesodeum. The results indicate that cathepsins B, D, and G and cathepsin H or aminopeptidase-like activities are present in the midgut of L. destructor. Among these activities, cathepsin D-like activity was identified for the first time in the gut of L. destructor. All proteases mentioned are produced in the mesodeal lumen and form the food bolus together with ingested food, afterward passing through the gut to be defecated. The method used enables the visualization of protease activities in the gut of transparent animals.
2. Sensitization and exposure to house dust and storage mites in high-altitude areas of ecuador
Rommel Valdivieso, Victor Iraola, Monica Estupiñán, Enrique Fernández-Caldas Ann Allergy Asthma Immunol. 2006 Oct;97(4):532-8. doi: 10.1016/S1081-1206(10)60946-5.
Background: Few studies have addressed exposure and sensitization to mite allergens in Andean countries. Objectives: To identify the main mite species in 3 locations at different altitudes in Ecuador and to verify skin test reactivity to various mite species in allergic individuals in Quito, Ecuador. Methods: Mattress dust samples were collected in Quito (2,800 m above sea level), Cuenca (2,500 m above sea level), and Guayaquil (sea level). Mite species present in the samples were isolated, identified, and counted. Der p 1 and Der f 1 levels were measured using monoclonal antibody-based enzyme immunoassays. Four hundred thirty-five patients in Quito diagnosed as having allergic rhinitis or asthma underwent skin testing with commercial extracts of Dermatophagoides pteronyssinus, Dermatophagoides farinae, Blomia tropicalis, Tyrophagus putrescentiae, and Lepidoglyphus destructor. In addition, Glycyphagus domesticus, Acarus siro, and Aleuroglyphus ovatus were tested in 362, 262, and 279 patients, respectively. Results: Twenty-one mite species were identified. Large populations of mites were detected above 2,500 m of altitude. All the dust samples contained detectable levels of Der p 1 or Der f 1. Positive skin prick test reactions to D. pteronyssinus, D. farinae, B. tropicalis, L. destructor, T. putrescentiae, A. ovatus, A. siro, and G. domesticus were obtained in 60.9%, 56.8%, 17.0%, 19.3%, 10.6%, 15.8%, 8.8%, and 11.0% of the patients, respectively. Conclusions: Most analyzed mattresses contained several species of mites. Mite allergen levels were high. This study confirms the importance of house dust and storage mite allergens in Ecuador in areas above 2,500 m of altitude, where humidity remains high year round.
3. The importance of starch and sucrose digestion in nutritive biology of synanthropic acaridid mites: alpha-amylases and alpha-glucosidases are suitable targets for inhibitor-based strategies of mite control
Tomas Erban, Michaela Erbanova, Marta Nesvorna, Jan Hubert Arch Insect Biochem Physiol. 2009 Jul;71(3):139-58. doi: 10.1002/arch.20312.
The adaptation of nine species of mites that infest stored products for starch utilization was tested by (1) enzymatic analysis using feces and whole mite extracts, (2) biotests, and (3) inhibition experiments. Acarus siro, Aleuroglyphus ovatus, and Tyroborus lini were associated with the starch-type substrates and maltose, with higher enzymatic activities observed in whole mite extracts. Lepidoglyphus destructor was associated with the same substrates but had higher activities in feces. Dermatophagoides farinae, Chortoglyphus arcuatus, and Caloglyphus redickorzevi were associated with sucrose. Tyrophagus putrescentiae and Carpoglyphus lactis had low or intermediate enzymatic activity on the tested substrates. Biotests on starch additive diets showed accelerated growth of species associated with the starch-type substrates. The inhibitor acarbose suppressed starch hydrolysis and growth of the mites. We suggest that the species with higher starch hydrolytic activity in feces were more tolerant to acarbose, and alpha-amylase and alpha-glucosidase of synanthropic mites are suitable targets for inhibitor-based strategies of mite control.
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