Z-Phe-Arg-AMC . HCl
目录号 : GA23894Z-Phe-Arg-AMC 是丝氨酸蛋白酶的底物,包括组织蛋白酶、激肽释放酶和纤溶酶。
Cas No.:70382-26-2
Sample solution is provided at 25 µL, 10mM.
Z-Phe-Arg-AMC . HCl is a substrate for serine proteases, including cathepsins, kallikrein and plasmin.Z-Phe-Arg-AMC is a substrate for serine proteases, including cathepsins, kallikrein and plasmin.Absorption/emission of substrate = 330/390 nm.
The Km and Vmax of the recombinant CsCPL-m towards Z-Phe-Arg-AMC were determined to be 5.71 × 10-6 M and 0.6 µM/min, respectively, at 37 °C and pH 5.5. The recombinant CsCPL-m could degrade BSA and gelatine, but could not degrade human hemoglobin and human immunoglobulin G[1].CatB and CatL activities in U2OS cell extracts were measured by fluorescent peptide conversion. The CatB/L-KO cells lacked CatB and CatL activity and were substantially resistant to EBOV GP-dependent entry[2].When measured the total endopeptidase activity of cysteine cathepsins in cell-free supernatant from both MPS and non-MPS samples, using Z-Phe-Arg-AMC as a broad-spectrum substrate, and E-64 as titration reagent. A 2.5-fold decrease of active cathepsins was measured in all MPS types compared with controls[3].
Plasma kallikrein activity was measured by the cleavage of the fluorometric substrate Z-Phe-Arg-AMC in plasma samples stimulated ex vivo with submaximal doses of dextran sulphate. Stimulated plasma kallikrein activity was significantly increased in both HAE-nl-C1INH and INHA subjects compared to non-swelling controls and histaminergic angioedema subjects. Using a threshold cut-off based on the normal controls[4].The Sphenophorus levis is one of the main pests of sugarcane in Brazil.Higher gene expression levels of Sl-CathL-CS occur in the midgut of 30-day old larvae. Sl-CathL-CS presented no protease activity, but Sl-CathL-mutSC hydrolyzed Z-Phe-Arg-AMC and was inhibited by a cysteine protease inhibitor E-64 (Ki = 38.52±1.20 µM), but not by the serine protease inhibitor PMSF[5].CPA in serum was determined with the spectrofluorometric technique using Z-Phe-Arg-AMC as a substrate. The highest CPA was found in breast cancer patients with a hereditary predisposition bearing BRCA1 gene mutations, and the lowest activity was found in patients who had a tumour surgically removed and before adjuvant therapy[6].When evaluated the capacity of negatively charged glycosaminoglycans to modulate the activity of catS. Chondroitin 4-sulfate (C4-S) impaired the collagenolytic activity and inhibited the peptidase activity (Z-Phe-Arg-AMC) of catS at pH 5.5, obeying a mixed-type mechanism[7].
References:
[1]: Ma C, Liang K, et,al. Identification and characteristics of a cathepsin L-like cysteine protease from Clonorchis sinensis. Parasitol Res. 2019 Mar;118(3):829-835. doi: 10.1007/s00436-019-06223-y. Epub 2019 Jan 28. PMID: 30689051.
[2]: Mittler E, Alkutkar T, et,al. Direct Intracellular Visualization of Ebola Virus-Receptor Interaction by In Situ Proximity Ligation. mBio. 2021 Jan 12;12(1):e03100-20. doi: 10.1128/mBio.03100-20. PMID: 33436438; PMCID: PMC7844541.
[3]: Chazeirat T, Denamur S, B et,al. The abnormal accumulation of heparan sulfate in patients with mucopolysaccharidosis prevents the elastolytic activity of cathepsin V. Carbohydr Polym. 2021 Feb 1;253:117261. doi: 10.1016/j.carbpol.2020.117261. Epub 2020 Oct 20. PMID: 33278943.
[4]: Lara-Marquez ML, Christiansen SC, et,al. Threshold-stimulated kallikrein activity distinguishes bradykinin- from histamine-mediated angioedema. Clin Exp Allergy. 2018 Nov;48(11):1429-1438. doi: 10.1111/cea.13219. Epub 2018 Aug 21. PMID: 29957871.
[5]: Shibao PYT, Ferro M, et,al. Identification and Functional Analysis of a Pseudo-Cysteine Protease from the Midgut Transcriptome of Sphenophorus levis. Int J Mol Sci. 2021 Oct 25;22(21):11476. doi: 10.3390/ijms222111476. PMID: 34768909; PMCID: PMC8583781.
[6]: Kilar E, Siewi¨½ski M, et,al. Differences in cysteine peptidases-like activity in sera of patients with breast cancer. Cancer Biomark. 2020;27(3):335-341. doi: 10.3233/CBM-190327. PMID: 31683457.
[7]: Sage J, MallÈvre F, et,al. Binding of chondroitin 4-sulfate to cathepsin S regulates its enzymatic activity. Biochemistry. 2013 Sep 17;52(37):6487-98. doi: 10.1021/bi400925g. Epub 2013 Sep 4. PMID: 23968158.
Z-Phe-Arg-AMC 是丝氨酸蛋白酶的底物,包括组织蛋白酶、激肽释放酶和纤溶酶。底物的吸收/发射 = 330/390 nm。
在 37 °C 和 pH 5.5 下,重组 CsCPL-m 对 Z-Phe-Arg-AMC 的 Km 和 Vmax 分别为 5.71 × 10-6 M 和 0.6 μM/min。重组CsCPL-m可降解BSA和明胶,但不能降解人血红蛋白和人免疫球蛋白G[1]。通过荧光肽转化法测定U2OS细胞提取物中CatB和CatL的活性。 CatB/L-KO 细胞缺乏 CatB 和 CatL 活性,对 EBOV GP 依赖性进入具有显着抗性[2]。和非 MPS 样品,使用 Z-Phe-Arg-AMC 作为广谱底物,E-64 作为滴定试剂。与对照组相比,所有 MPS 类型的活性组织蛋白酶减少了 2.5 倍[3]。
血浆激肽释放酶活性是通过在用亚最大剂量的硫酸葡聚糖离体刺激的血浆样品中荧光底物 Z-Phe-Arg-AMC 的裂解来测量的。与非肿胀对照和组胺能血管性水肿受试者相比,在 HAE-nl-ClINH 和 INHA 受试者中受刺激的血浆激肽释放酶活性显着增加。使用基于正常对照的阈值截断[4]。Sphenophorus levis 是巴西甘蔗的主要害虫之一。Sl-CathL-CS 的基因表达水平较高发生在中肠30 天大的幼虫。 Sl-CathL-CS 没有蛋白酶活性,但 Sl-CathL-mutSC 水解 Z-Phe-Arg-AMC 并被半胱氨酸蛋白酶抑制剂 E-64 (Ki = 38.52±1.20 μM) 抑制,但不受丝氨酸蛋白酶抑制以Z-Phe-Arg-AMC为底物,采用荧光分光光度法测定血清中的抑制剂PMSF[5].CPA。 CPA 最高的是具有遗传易感性的 BRCA1 基因突变的乳腺癌患者,而最低的活性是在手术切除肿瘤和辅助治疗前的患者[6]。带负电荷的糖胺聚糖调节 catS 活性的能力。 4-硫酸软骨素 (C4-S) 在 pH 5.5 时削弱了 catS 的胶原蛋白溶解活性并抑制了肽酶活性 (Z-Phe-Arg-AMC),遵循混合型机制[7]。
Kinase experiment [1]: | |
Preparation Method |
For Km determinations, Z-Phe-Arg-AMC.HCl was used as a substrate at different concentrations (4.0, 8.0, 12.0, 16.0, 20.0, 24.0, 28.0, 32.0, 36.0, 40.0, and 44.0 μM). The fluorescence intensity of AMC was measured continuously every 3 min. |
Reaction Conditions |
4.0-44.0 μM Z-Phe-Arg-AMC.HCl incubat with rCsCPL-m |
Applications |
Rcscpl-m showed protease activity against Z-Phe-Arg-AMC.HCl, The Km and Vmax of RCSCPL-M for Z-Phe-Arg-AMC.HCl were determined to be 5.71 × 10-6 M and 0.6 μm /min at 37 °C and pH 5.5. |
Cell experiment [2]: | |
Cell lines |
U2OS cell |
Preparation Method |
Cell lysates were mixed with reaction buffer, incubated with the fluorogenic peptide substrate Z-Phe-Arg-AMC.HCl (150 uM), and measured at a fluorometer following a 0- to 30-min incubation |
Reaction Conditions |
Z-Phe-Arg-AMC.HCl (150 uM) for 0-30min |
Applications |
CatB and CatL activities in U2OS cell extracts were measured by fluorescent peptide conversion( Z-Phe-Arg-AMC.HCl). To further investigate the cysteine protease-dependent EBOV entry mechanism, CatB/L-knockout (KO) U2OS cell lines by CRISPR/Cas9 genome engineering has been generated.The CatB/L-KO cells lacked CatB and CatL activity and were substantially resistant to EBOV GP-dependent entry. |
References: [1]. Ma C, Liang K, et,al. Identification and characteristics of a cathepsin L-like cysteine protease from Clonorchis sinensis. Parasitol Res. 2019 Mar;118(3):829-835. doi: 10.1007/s00436-019-06223-y. Epub 2019 Jan 28. PMID: 30689051. [2]. Mittler E, Alkutkar T, et,al. Direct Intracellular Visualization of Ebola Virus-Receptor Interaction by In Situ Proximity Ligation. mBio. 2021 Jan 12;12(1):e03100-20. doi: 10.1128/mBio.03100-20. PMID: 33436438; PMCID: PMC7844541. |
Cas No. | 70382-26-2 | SDF | |
分子式 | C33H36N6O6 · HCl | 分子量 | 649.15 |
溶解度 | Soluble in DMSO | 储存条件 | Store at -20°C |
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Shipping Condition | 评估样品解决方案:配备蓝冰进行发货。所有其他可用尺寸:配备RT,或根据请求配备蓝冰。 |
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1 mg | 5 mg | 10 mg | |
1 mM | 1.5405 mL | 7.7024 mL | 15.4048 mL |
5 mM | 0.3081 mL | 1.5405 mL | 3.081 mL |
10 mM | 0.154 mL | 0.7702 mL | 1.5405 mL |
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% DMSO % % Tween 80 % saline | ||||||||||
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2.
一定要按照顺序依次将溶剂加入,进行下一步操作之前必须保证上一步操作得到的是澄清的溶液,可采用涡旋、超声或水浴加热等物理方法助溶。
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Quality Control & SDS
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- Purity: >98.50%
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