Ac-ANW-AMC is a fluorescent substrate for the β5i (LMP7/PSMB8) subunit of the 20S immunoproteasome, with a maximum excitation wavelength of λEx=345nm and a maximum emission wavelength of λEm=445nm[1, 2]. When a specific enzyme acts on Ac-ANW-AMC, it releases the fluorophore 7-amido-4-methylcoumarin (AMC). The activity of the enzyme can be quantitatively analyzed by measuring the fluorescence intensity of AMC[3]. The fluorescence signal of Ac-ANW-AMC can be effectively eliminated by the i-proteasome inhibitor ONX-0914[4].
References:
[1] Winter M B, La Greca F, Arastu-Kapur S, et al. Immunoproteasome functions explained by divergence in cleavage specificity and regulation[J]. Elife, 2017, 6: e27364.
[2] Erokhov P A, Kulikov A M, Karpova Y D, et al. Proteasomes in patient rectal cancer and different intestine locations: where does proteasome pool change?[J]. Cancers, 2021, 13(5): 1108.
[3] Mustafa M S, El-Abadelah M M, Mubarak M S, et al. Synthesis and fluorogenic properties of some 1-(coumarin-7-yl)-4, 5-dihydro-1, 2, 4-triazin-6 (1H)-ones[J]. International Journal of Chemistry, 2011, 3(4): 89.
[4] Kim S, Park S H, Choi W H, et al. Evaluation of Immunoproteasome-Specific Proteolytic Activity Using Fluorogenic Peptide Substrates[J]. Immune Network, 2022, 22(3).
Ac-ANW-AMC是20S免疫蛋白酶体的β5i（LMP7/PSMB8）亚基的荧光底物，最大激发波长λEx=345nm, 最大发射波长λEm=445nm[1, 2]。当特定酶作用于Ac-ANW-AMC时，会释放出荧光团7-amido-4-methylcoumarin (AMC)，通过测量AMC的荧光强度可以定量分析酶的活性[3]。Ac-ANW-AMC的荧光信号能够被i-蛋白酶体抑制剂ONX-0914有效消除[4]。