Papain (≥800u/mg)
(Synonyms: 木瓜酶) 目录号 : GC33829
木瓜蛋白酶是一种具有广泛特异性的半胱氨酸蛋白酶，可切割碱性氨基酸、亮氨酸或甘氨酸的肽键。
Cas No.:9001-73-4
Sample solution is provided at 25 µL, 10mM.
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Papain is a cysteine protease with wide specificity, cleaving peptide bonds of basic amino acids, leucine, or glycine. It also hydrolyzes esters and amides.
Papain did not induce proteolysis or cell detachment of HUVEC in a concentration range between 0 and 25 μg/mL. Four hours treatment with 10 μg/mL papain resulted in a reduced susceptibility of endothelial cells to activation by VEGF as determined by phosphorylation levels of Akt, MEK1/2, SAPK/JNK. Papain exerted a distinct inhibitory effect on cell growth, cell migration and tube formation with inhibition of tube formation detectable at concentrations as low as 1 μg/mL[4]. Papain Activates Human Mast Cells to Release Proinflammatory Mediators via its Enzymatic Activity, papain is a direct activator of both human skin MCs (connective tissue) and CBMCs (mucosal type) via its protease activity and partly via PAR-2 [5]. While the affinity to cGMP, I-V relation, channel kinetics, and single-channel current amplitude were almost the same in both papain-treated and the control preparations, the total current flowing through the patch membrane of the papain-treated cells was reduced to about 20% of the control, suggesting that papain reduced the density of cGMP-activated channels[2]. A significant increase in cell viability when papain was used for RPC isolation[3].
Papain is a cysteine protease allergen with inherent adjuvant activity, induces potent IL-4 expression by T cells in the popliteal lymph nodes of mice following footpad immunization. B cells subsequently regulate the adaptive immune response by enhancing ICOS expression on CD4(+) T cells and amplifying Th2 and follicular helper T cell induction. Ab blockade of ICOS ligand, expressed by popliteal lymph node B cells, but not dendritic cells, at the peak of the response inhibits IL-4 responses in wild-type mice but not B cell-deficient mice. Thus, B cells play a critical role in amplifying adjuvant-dependent Th2 polarization following noncanonical acquisition and internalization of the cysteine protease papain[1]. purified papain from C. papaya were effective against S. venezuelensis eggs and larvae in vitro, suggesting their potential use as an alternative treatment for strongyloidiasis[7]. Rapidly progressing joint disease can be produced in the rabbit by intra-articular injection of concentrated papain[9]. Enzymatically active papain preferentially induces an IgG1 response and results in mast cell degranulation, both features typical of an allergic reaction in mice[8].
References:
[1]. Dwyer DF, Woodruff MC, et,al. B cells regulate CD4+ T cell responses to papain following B cell receptor-independent papain uptake. J Immunol. 2014 Jul 15;193(2):529-39. doi: 10.4049/jimmunol.1303247. Epub 2014 Jun 13. PMID: 24928991; PMCID: PMC4203309.
[2]. Shen J, Watanabe S, et,al. Cell dissociation with papain reduces the density of cGMP-activated channels of the retinal rod. Jpn J Physiol. 1995;45(1):151-64. doi: 10.2170/jjphysiol.45.151. PMID: 7544417.
[3]. Scruggs BA, Jiao C, et,al. Optimizing Donor Cellular Dissociation and Subretinal Injection Parameters for Stem Cell-Based Treatments. Stem Cells Transl Med. 2019 Aug;8(8):797-809. doi: 10.1002/sctm.18-0210. Epub 2019 Apr 19. PMID: 31004408; PMCID: PMC6646699.
[4]. Mohr T, Desser L. Plant proteolytic enzyme papain abrogates angiogenic activation of human umbilical vein endothelial cells (HUVEC) in vitro. BMC Complement Altern Med. 2013 Sep 21;13:231. doi: 10.1186/1472-6882-13-231. PMID: 24053149; PMCID: PMC3849051.
[5]. Seaf M, Ben-Zimra M, et,al. Papain Activates Human Mast Cells to Release Proinflammatory Mediators via its Enzymatic Activity. J Invest Dermatol. 2016 Jul;136(7):1523-1525. doi: 10.1016/j.jid.2016.03.030. Epub 2016 Apr 7. PMID: 27060447.
[6]. Moraes D, Levenhagen MA, et,al. In vitro efficacy of latex and purified papain from Carica papaya against Strongyloides venezuelensis eggs and larvae. Rev Inst Med Trop Sao Paulo. 2017 Apr 3;59:e7. doi: 10.1590/S1678-9946201759007. PMID: 28380118; PMCID: PMC5441158.
[7]. Moraes D, Levenhagen MA, et,al. In vitro efficacy of latex and purified papain from Carica papaya against Strongyloides venezuelensis eggs and larvae. Rev Inst Med Trop Sao Paulo. 2017 Apr 3;59:e7. doi: 10.1590/S1678-9946201759007. PMID: 28380118; PMCID: PMC5441158.
[8]. Chambers L, Brown A, et,al. Enzymatically active papain preferentially induces an allergic response in mice. Biochem Biophys Res Commun. 1998 Dec 30;253(3):837-40. doi: 10.1006/bbrc.1998.9862. PMID: 9918815.
[9]. Havdrup T, Telhag H. Papain-induced changes in the knee joints of adult rabbits. Acta Orthop Scand. 1977;48(2):143-9. doi: 10.3109/17453677708985125. PMID: 868495.
木瓜蛋白酶是一种具有广泛特异性的半胱氨酸蛋白酶，可切割碱性氨基酸、亮氨酸或甘氨酸的肽键。它还水解酯和酰胺。
在 0 到 25 μg/mL 的浓度范围内，木瓜蛋白酶不会诱导 HUVEC 的蛋白水解或细胞分离。用 10 μg/mL 木瓜蛋白酶处理 4 小时导致内皮细胞对 VEGF 激活的敏感性降低，这由 Akt、MEK1/2、SAPK/JNK 的磷酸化水平确定。木瓜蛋白酶对细胞生长、细胞迁移和管形成具有明显的抑制作用，在低至 1 μg/mL 的浓度下可检测到抑制管形成[4]。木瓜蛋白酶通过其酶活性激活人类肥大细胞释放促炎介质，木瓜蛋白酶通过其蛋白酶活性和部分通过 PAR-2 [5] 直接激活人类皮肤 MC（结缔组织）和 CBMC（粘膜类型） 。虽然木瓜蛋白酶处理和对照制剂对 cGMP 的亲和力、I-V 关系、通道动力学和单通道电流幅度几乎相同，但流过木瓜蛋白酶处理细胞贴片膜的总电流减少至约对照的 20%，表明木瓜蛋白酶降低了 cGMP 激活通道的密度[2]。当使用木瓜蛋白酶分离 RPC 时，细胞活力显着增加[3]。
木瓜蛋白酶是一种具有固有佐剂活性的半胱氨酸蛋白酶过敏原，在足垫免疫后诱导小鼠腘淋巴结中 T 细胞的强效 IL-4 表达。 B 细胞随后通过增强 ICOS 在 CD4(+) T 细胞上的表达和放大 Th2 和滤泡辅助性 T 细胞诱导来调节适应性免疫反应。由腘淋巴结 B 细胞而非树突细胞表达的 ICOS 配体的 Ab 阻断在反应的峰值抑制野生型小鼠而非 B 细胞缺陷小鼠的 IL-4 反应。因此，在半胱氨酸蛋白酶木瓜蛋白酶[1] 的非典型获得和内化后，B 细胞在放大佐剂依赖性 Th2 极化中起着关键作用。来自 C. papaya 的纯化木瓜蛋白酶在体外对 S. venezuelensis 卵和幼虫有效，表明它们可用作类圆线虫病的替代疗法[7]。兔关节内注射浓缩木瓜蛋白酶[9]可引起快速进展的关节病。具有酶促活性的木瓜蛋白酶优先诱导 IgG1 反应并导致肥大细胞脱颗粒，这两个特征都是小鼠过敏反应的典型特征[8]。
| Cell experiment [1]: | |
|
Cell lines |
HUVEC cell |
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Preparation Method |
Cells seeded into 12 well plates and incubated with EGM to allow for the formation of monolayers. After 24 hours, EGM was replaced with EBM containing 2.5% FCS, 10 ng/mL VEGF, and 10 µg/mL papain. After a further 10 hours incubation period, cells were stained with 2 µM Calcein and pictures were taken at 4 fold magnification using a Nikon Eclipse Ti, the FITC filter set of the instrument and a Nikon Digital Sight DS-Fi1C camera. |
|
Reaction Conditions |
10 µg/mL papain for 10 hours |
|
Applications |
Papain did not induce proteolysis or cell detachment of HUVEC in a concentration range between 0 and 25 µg/mL. Four hours treatment with 10 µg/mL papain resulted in a reduced susceptibility of endothelial cells to activation by VEGF as determined by phosphorylation levels of Akt, MEK1/2, SAPK/JNK. Papain exerted a distinct inhibitory effect on cell growth, cell migration and tube formation with inhibition of tube formation detectable at concentrations as low as 1 µg/mL. |
| Animal experiment [2]: | |
|
Animal models |
C57BL/6 mice |
|
Preparation Method |
C57BL/6 mice were given naive B cells by i.v. injection 24 h prior to imaging. Recipient mice were anesthetized with 2% isoflurane and prepared on a surgical board for PLN live imaging. Exposed PLNs were monitored for physiological temperature maintenance and imaged using a Zeiss Bio-Rad Radiance 2100 Multiphoton microscope.AF488-labeled papain was injected in the footpad at time point 0, and serial 150-µm depth captures (3-µm steps) were taken every 2 min for 60 min following papain injection. |
|
Dosage form |
For intravital microscopy, mice were injected with 10 µl papain at a concentration of 10 mg/ml. |
|
Applications |
Papain, a cysteine protease allergen with inherent adjuvant activity, induces potent IL-4 expression by T cells in the popliteal lymph nodes of mice following footpad immunization. B cells subsequently regulate the adaptive immune response by enhancing ICOS expression on CD4(+) T cells and amplifying Th2 and follicular helper T cell induction. Ab blockade of ICOS ligand, expressed by popliteal lymph node B cells, but not dendritic cells, at the peak of the response inhibits IL-4 responses in wild-type mice but not B cell-deficient mice. Thus, B cells play a critical role in amplifying adjuvant-dependent Th2 polarization following noncanonical acquisition and internalization of the cysteine protease papain. |
|
References: [1].Mohr T, Desser L. Plant proteolytic enzyme papain abrogates angiogenic activation of human umbilical vein endothelial cells (HUVEC) in vitro. BMC Complement Altern Med. 2013 Sep 21;13:231. doi: 10.1186/1472-6882-13-231. PMID: 24053149; PMCID: PMC3849051. |
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| Cas No. | 9001-73-4 | SDF | |
| 别名 | 木瓜酶 | ||
| Canonical SMILES | [Papain] | ||
| 分子式 | 分子量 | ||
| 溶解度 | Water : 50 mg/mL (Need ultrasonic) ;DMSO : 5 mg/mL (Need ultrasonic) | 储存条件 | Store at 2-8°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。 | ||
| 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:
- Biological Activity: 803u/mg
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet