Z-Ala-Ala-Leu-pNA CAS NO.61043-33-2

Z-Ala-Ala-Leu-pNA (CAS NO. 61043-33-2) is a synthetic, chromogenic tripeptide substrate specifically designed for the detection and quantification of chymotrypsin-like serine protease activity. This research-grade compound features a carboxybenzyl (Z) protecting group, an Ala-Ala-Leu recognition sequence, and a para-nitroaniline (pNA) leaving group, enabling sensitive and direct spectrophotometric measurement of enzymatic hydrolysis.

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TECHNICAL SPECIFICATIONS

Item	Specification
Product Name	Z-Ala-Ala-Leu-pNA
CAS NO.	61043-33-2
Appearance	White to off-white lyophilized powder
Molecular Formula	C₂₆H₃₃N₅O₇
Molecular Weight	527.6 g/mol
Purity (RP-HPLC)	≥98%
Solubility	Soluble in water, DMSO, and aqueous buffers
Endotoxin	≤1.0 EU/mg
Biological Activity	Chromogenic substrate for serine proteases (e.g., Subtilisin A), used to measure protease activity in vitro
Storage Conditions	Store at -20°C (sealed, protected from light and moisture); short-term storage at +4°C is acceptable.
Stock Location	Stock in USA
MOQ	1g

1.MECHANISM OF ACTION

This substrate acts through a targeted protease cleavage-release mechanism. The Ala-Ala-Leu sequence is specifically recognized and cleaved at the carboxyl side of the leucine (Leu) residue by chymotrypsin-like enzymes, which prefer large hydrophobic amino acids at the P1 position. Upon cleavage by an active protease (e.g., chymotrypsin, cathepsin G, neutrophil elastase to a lesser extent), the para-nitroaniline (pNA) moiety is liberated. Free pNA absorbs light strongly at 405-410 nm, producing a yellow color. The rate of increase in absorbance at this wavelength is directly proportional to the enzyme's activity.

2.PRODUCT INDICATIONS

This product is indicated for biochemical and pharmacological research involving chymotrypsin-like proteases. Key applications include: assaying the activity of purified chymotrypsin, cathepsin G, or related serine proteases; screening for and characterizing inhibitors of these enzymes in drug discovery (e.g., for inflammatory or lung diseases); measuring chymotrypsin-like activity in complex biological samples like tissue homogenates or cell lysates; and as a standard substrate in enzyme kinetics studies to determine kinetic parameters (Km, Vmax).

3.CLINICAL EFFICACY OF PRODUCT

Z-Ala-Ala-Leu-pNA is a diagnostic and research reagent, not a therapeutic. Its clinical efficacy is demonstrated through its indispensable role in biomarker research and drug development. It is used in: 1) Disease Research: To measure elevated levels of chymotrypsin-like proteases (e.g., in neutrophil-dominated inflammation, pancreatitis, or certain cancers) as potential biomarkers. 2) Therapeutic Monitoring: In vitro assays to monitor the efficacy of protease inhibitor drugs. 3) Drug Discovery: As a primary screening tool to identify and optimize novel inhibitors targeting chymotrypsin, cathepsin G, or the proteasome's chymotrypsin-like activity, supporting the development of treatments for conditions like emphysema, cystic fibrosis, and multiple myeloma.

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