Leupeptin, Microbial
(Synonyms: 亮肽素) 目录号 : GC10027
Leupeptin, Microbial 是一种广谱丝氨酸和半胱氨酸蛋白酶抑制剂。
Cas No.:103476-89-7
Sample solution is provided at 25 µL, 10mM.
Leupeptin, Microbial is a broad-spectrum serine and cysteine protease inhibitor [1]. Leupeptin, Microbial is a reversible protease inhibitor, It acts on bovine trypsin, human plasminase, bovine splenic cathepsin B, and recombinant human caltrypsin with Ki values of 35 nM, 3.4 μM, 6 nM, and 72 nM[7].
The protease inhibitor Leupeptin, Microbial prevented multiplication of the human coronavirus strain 229E in cultures of MRC-C cells [3]. Leupeptin, Microbial-mediated inhibition of trypsin-like proteases maintains substrate mycelium development, whereas proteolytic degradation of Leupeptin, Microbial in stationary phase cultures derepresses the trypsin-like proteases, leading to the digestion of substrate mycelium and promotion of aerial mycelium formation [2]. As the calpain inhibitor Leupeptin, Microbial primarily protected hair cells from neomycin[5]. The expression of hepatitis B surface antigen (HBsAg) from Leupeptin recovered up to 50% of the cell suspension culture [6].
Leupeptin, Microbial was well tolerated in mice. Leupeptin, Microbial significantly increased LC3b-II in a dose-dependent way in the total tissue extract and lysosomal enrichment portion (LE portion). At the level of electron microscopy (EM), Leupeptin, Microbial induced the accumulation of electron-dense vesicle structures [4].
References:
[1]. Aoyagi T, Miyata S, et,al. Biological activities of leupeptins. J Antibiot (Tokyo). 1969 Nov;22(11):558-68. doi: 10.7164/antibiotics.22.558. PMID: 4243683.
[2]. Kim IS, Kim YB, et,al. Characterization of the leupeptin-inactivating enzyme from Streptomyces exfoliatus SMF13 which produces leupeptin. Biochem J. 1998 Apr 15;331 ( Pt 2)(Pt 2):539-45. doi: 10.1042/bj3310539. PMID: 9531495; PMCID: PMC1219386.
[3]. Appleyard G, Tisdale M. Inhibition of the growth of human coronavirus 229E by leupeptin. J Gen Virol. 1985 Feb;66 ( Pt 2):363-6. doi: 10.1099/0022-1317-66-2-363. PMID: 3968542.
[4]. Haspel J, Shaik RS, et,al. Characterization of macroautophagic flux in vivo using a leupeptin-based assay. Autophagy. 2011 Jun;7(6):629-42. doi: 10.4161/auto.7.6.15100. Epub 2011 Jun 1. PMID: 21460622; PMCID: PMC3127049.
[5]. Aoyagi T, Miyata S, et,al. Biological activities of leupeptins. J Antibiot (Tokyo). 1969 Nov;22(11):558-68. doi: 10.7164/antibiotics.22.558. PMID: 4243683.
[6]. Coffin AB, Williamson KL, et,al. Profiling drug-induced cell death pathways in the zebrafish lateral line. Apoptosis. 2013 Apr;18(4):393-408. doi: 10.1007/s10495-013-0816-8. PMID: 23413197; PMCID: PMC3627356.
[7]. Ganapathi TR, Sunil Kumar GB, et,al. Analysis of the limitations of hepatitis B surface antigen expression in soybean cell suspension cultures. Plant Cell Rep. 2007 Sep;26(9):1575-84. doi: 10.1007/s00299-007-0379-7. Epub 2007 May 30. PMID: 17534624.
Leupeptin, Microbial 是一种广谱丝氨酸和半胱氨酸蛋白酶抑制剂[1]。 Leupeptin, Microbial 是一种可逆的蛋白酶抑制剂，它作用于牛胰蛋白酶、人纤溶酶、牛脾脏组织蛋白酶 B 和重组人钙蛋白酶，Ki 值为 35 nM、3.4 μM、6 nM 和 72 nM[7].
蛋白酶抑制剂 Leupeptin Microbial 阻止了 MRC-C 细胞培养物中人类冠状病毒株 229E 的增殖[3]。亮肽素，微生物介导的胰蛋白酶样蛋白酶抑制维持底物菌丝体发育，而亮肽素的蛋白水解降解，微生物在稳定期培养物中去抑制胰蛋白酶样蛋白酶，导致底物菌丝体的消化和气生菌丝体形成的促进 [2].作为钙蛋白酶抑制剂 Leupeptin，Microbial 主要保护毛细胞免受新霉素的侵害[5]。 Leupeptin的乙型肝炎表面抗原（HBsAg）的表达在细胞悬浮培养物中恢复了50%[6]。
Leupeptin, Microbial 在小鼠中具有良好的耐受性。 Leupeptin, Microbial 在总组织提取物和溶酶体富集部分（LE 部分）中以剂量依赖性方式显着增加 LC3b-II。在电子显微镜（EM）水平，亮肽素、微生物诱导电子致密囊泡结构的积累[4]。
| Leupeptin analysis [1]: | |
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Preparation Method |
The concentration of Leupeptin, Microbial was calculated by enzyme inhibition assay using 50 pg of papain and 0.2 mM of N-benzoylL-arginine ethyl ester as the target protease and papain substrate, respectively. Enzyme reactions were carried out at 25 ℃ and pH 6.2. |
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Applications |
Leupeptin, Microbial is a leucine-specific protease and a metalloprotease. |
| Cell experiment [2]: | |
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Cell lines |
MRC-C cell |
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Preparation Method |
To investigate the effect of Leupeptin, Microbial on virus yield, MRC-C cultures in 35 mm dishes were infected by adsorption of virus from 0T ml concentrated suspension of HCV 229E for 1 h at 36 ℃. The cultures were washed twice and overlaid with 2 ml Eagle’s MEM plus 0.2% bovine serum albumin and 20 mm-HEPES buffer pH 7.4. Leupeptin, Microbial at a range of concentrations was added to the virus inoculum, to the maintenance medium and also to maintenance medium used to treat the cultures for 1 h before infection. After 24 h at 36 ℃ in air, the cells were scraped into the medium and disrupted by sonication. |
|
Reaction Conditions |
0.4 to l00 µg/ml for 25 h（1 h before infection to 24 h after infection） |
|
Applications |
The protease inhibitor Leupeptin, Microbial prevented multiplication of the human coronavirus strain 229E in cultures of MRC-C cells. |
| Animal experiment [3]: | |
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Animal models |
C57BL/6NCrl male mouse |
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Preparation Method |
Mice received i.p. injections of 0.5 ml sterile Phosphate Buffered Saline or 0.5 ml PBS containing 9–40 mg/kg Leupeptin, Microbial hemisulfate. |
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Dosage form |
0, 9, 18 36 and 40 mg/kg Leupeptin, Microbial; Intraperitoneal injection |
|
Applications |
Rate of LC3b accumulation after Leupeptin, Microbial treatment was greatest in the liver and lowest in spleen. |
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References: [1]. Kim IS, Kim YB, et,al. Characterization of the leupeptin-inactivating enzyme from Streptomyces exfoliatus SMF13 which produces leupeptin. Biochem J. 1998 Apr 15;331 (Pt 2)(Pt 2):539-45. doi: 10.1042/bj3310539. PMID: 9531495; PMCID: PMC1219386. [2]. Appleyard G, Tisdale M. Inhibition of the growth of human coronavirus 229E by leupeptin. J Gen Virol. 1985 Feb;66 (Pt 2):363-6. doi: 10.1099/0022-1317-66-2-363. PMID: 3968542. [3]. Haspel J, Shaik RS, et,al. Characterization of macroautophagic flux in vivo using a leupeptin-based assay. Autophagy. 2011 Jun;7(6):629-42. doi: 10.4161/auto.7.6.15100. Epub 2011 Jun 1. PMID: 21460622; PMCID: PMC3127049. |
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| Cas No. | 103476-89-7 | SDF | |
| 别名 | 亮肽素 | ||
| 化学名 | 2-acetamido-N-(1-((5-((diaminomethylene)amino)-1-oxopentan-2-yl)amino)-4-methyl-1-oxopentan-2-yl)-4-methylpentanamide | ||
| Canonical SMILES | O=C(C(C([H])([H])C(C([H])([H])[H])([H])C([H])([H])[H])([H])N([H])C(C([H])([H])[H])=O)N([H])C(C(N([H])C(C([H])=O)([H])C([H])([H])C([H])([H])C([H])([H])/N=C(N([H])[H])/N([H])[H])=O)([H])C([H])([H])C(C([H])([H])[H])([H])C([H])([H])[H] | ||
| 分子式 | C20H38N6O4.1/2H2SO4 | 分子量 | 493.6 |
| 溶解度 | ≥ 24.7mg/mL in DMSO， ≥ 20mg/mL in Water | 储存条件 | Store at -20°C |
| General tips | 请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。 储备液的保存方式和期限：-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。 为了提高溶解度，请将管子加热至37℃，然后在超声波浴中震荡一段时间。 |
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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。 | ||
| 制备储备液 | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 2.0259 mL | 10.1297 mL | 20.2593 mL |
| 5 mM | 0.4052 mL | 2.0259 mL | 4.0519 mL |
| 10 mM | 0.2026 mL | 1.013 mL | 2.0259 mL |
| 第一步：请输入基本实验信息（考虑到实验过程中的损耗，建议多配一只动物的药量） | ||||||||||
| 给药剂量 | mg/kg |  |
动物平均体重 | g | |
每只动物给药体积 | ul | |
动物数量 | 只 |
| 第二步：请输入动物体内配方组成（配方适用于不溶于水的药物；不同批次药物配方比例不同，请联系GLPBIO为您提供正确的澄清溶液配方） | ||||||||||
| % DMSO % % Tween 80 % saline | ||||||||||
| 计算重置 | ||||||||||
计算结果:
工作液浓度： mg/ml；
DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL，
体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL saline，混匀澄清。
1. 首先保证母液是澄清的；
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。
3. 以上所有助溶剂都可在 GlpBio 网站选购。
Quality Control & SDS
- View current batch:
- Purity: >98.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet