Z-Ala-Ala-Asn-AMC (Cbz-Ala-Ala-Asn-AMC) is a synthetic fluorogenic peptide substrate that releases highly fluorescent AMC upon cleavage by proteases (Ex/Em: 380nm/430nm), which is widely used for the detection of asparaginyl endopeptidase (Legumain) activity[1,2,3]. Legumain is often overexpressed in tumors[4], and Z-Ala-Ala-Asn-AMC serves as a substrate suitable for detecting and quantifying legumain activity in biological samples, high-throughput inhibitor screening, and enzyme kinetics studies[5,6,7].
References:
[1] ZHAO T, LI Z, GUO Z, et al. Functional recombinant human Legumain protein expression in Pichia pastoris to enable screening for Legumain small molecule inhibitors[J]. Protein Expression and Purification, 2018, 150: 12-16.
[2] POREBA M. Recent advances in the development of legumain-selective chemical probes and peptide prodrugs[J]. Biological Chemistry, 2019, 400(12): 1529-1550.
[3] PICKERING A M, DAVIES K J A. A simple fluorescence labeling method for studies of protein oxidation, protein modification, and proteolysis[J]. Free Radical Biology and Medicine, 2012, 52(2): 239-246.
[4] LIU C, SUN C, HUANG H, et al. Overexpression of legumain in tumors is significant for invasion/metastasis and a candidate enzymatic target for prodrug therapy[J]. Cancer Research, 2003, 63(11): 2957-2964.
[5] ZHAO T, LIU Y, HAO Y, et al. Esomeprazole inhibits the lysosomal cysteine protease legumain to prevent cancer metastasis[J]. Investigational New Drugs, 2021, 39(2): 337-347.
[6] POREBA M, SOLBERG R, RUT W, et al. Counter selection substrate library strategy for developing specific protease substrates and probes[J]. Cell Chemical Biology, 2016, 23(8): 1023-1035.
[7] GRUNCLOVA L, HORN M, VANCOVÁ M, et al. Two secreted cystatins of the soft tick Ornithodoros moubata: differential expression pattern and inhibitory specificity[J]. Biological Chemistry, 2006, 387(12).
Z-Ala-Ala-Asn-AMC（Cbz-Ala-Ala-Asn-AMC）是一种在被蛋白酶切割后可释放高荧光强度AMC的合成荧光肽底物（Ex/Em: 380nm/430nm），被广泛用于天冬酰胺内肽酶（Legumain）活性的测定[1,2,3]。Legumain是一种在肿瘤中常过度表达的酶[4]，Z-Ala-Ala-Asn-AMC作为底物适用于生物样本中豆苷活性的检测定量、高通量抑制剂筛选和酶动力学研究等领域[5,6,7]。