Pepstatin A
(Synonyms: 抑肽素; Pepstatin A) 目录号 : GC11974
Pepstatin A is an orally active inhibitor of aspartic proteases, which is produced by actinomycetes.
Cas No.:26305-03-3
Sample solution is provided at 25 µL, 10mM.
Quality Control & SDS
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- Purity: >98.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
| Cell experiment [1]: | |
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Cell lines |
Osteoclasts |
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Preparation Method |
Pepstatin A was added at 15-120 µM to the growth medium. |
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Reaction Conditions |
0-120 µM |
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Applications |
Pepstatin A suppressed the formation of TRAP-positive multinuclear cells in a dose-dependent manner. |
| Animal experiment [2]: | |
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Animal models |
Ligated Shay rats |
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Dosage form |
0.5-50 mg/kg, p.o. |
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Applications |
After using Pepstatin A, free sialic acid and bound sialic acid were decreased in the gastric juice of the Shay rats. |
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References: [1]. Yoshida H, Okamoto K, et,al. Pepstatin A, an aspartic proteinase inhibitor, suppresses RANKL-induced osteoclast differentiation. J Biochem. 2006 Mar;139(3):583-90. doi: 10.1093/jb/mvj066. PMID: 16567424. |
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Pepstatin A is an orally active inhibitor of aspartic proteases, which is produced by actinomycetes. It exhibits inhibitory activity with IC50 values of 4.5 nM, 6.2 nM, 150 nM, 290 nM, 520 nM, and 260 nM against hemoglobin-pepsin, hemoglobin-proctase, casein-pepsin, casein-proctase, casein-acid protease, and hemoglobin-acid protease, respectively. Additionally, pepstatin has been found to inhibit HIV protease [1-3].
Pepstatin A(0-120 µM) suppressed receptor activator of NF-κB ligand (RANKL)-induced osteoclast differentiation[4]. Pepstatin A(0.1-0.3 µM) caused a reproducible, concentration-related increase in the extracellular acidification rate in two microglial cell lines, Ra2 and 6-3[5]. Pepstatin A significantly hindered influenza virus replication, probably by modulating host cell autophagic/apoptotic responses[6].
Pepstatin (0.5-50 mg/kg; p.o.) suppresses stomach ulceration of the pylorus in ligated Shay rats[1].
胃抑素Pepstatin A是由放线菌类产生的一种特异性的、具有口服活性的天冬氨酸蛋白酶 (aspartic proteases) 抑制剂,能够抑制 hemoglobin-pepsin、hemoglobin-proctase、casein-pepsin、casein-proctase、casein-acid protease 和 hemoglobin-acid protease 的活性,IC50 值分别为 4.5 nM、6.2 nM、150 nM、290 nM、520 nM 和 260 nM；Pepstatin A可抑制 HIV protease 的活性[1-3].
Pepstatin A (0-120 µM) 抑制了受体活化因子NF-κB配体(RANKL)-诱导的破骨细胞分化[4]。Pepstatin A (0.1-0.3 µM) 可以在两个微胶质细胞系Ra2和6-3中引起浓度相关的可重复的细胞外酸化率增加[5]。Pepstatin A 显著阻碍了流感病毒的复制,可能是通过调节宿主细胞自噬/凋亡反应发挥作用[6]。
Pepstatin A (0.5-50 mg/kg; p.o.)对结扎的谢氏大鼠幽门溃疡有抑制作用[1]。
References:
[1]. Yoshida H, Okamoto K, et,al. Pepstatin A, an aspartic proteinase inhibitor, suppresses RANKL-induced osteoclast differentiation. J Biochem. 2006 Mar;139(3):583-90. doi: 10.1093/jb/mvj066. PMID: 16567424.
[2]. Seelmeier S, Schmidt H, et,al. Human immunodeficiency virus has an aspartic-type protease that can be inhibited by pepstatin A. Proc Natl Acad Sci U S A. 1988 Sep;85(18):6612-6. doi: 10.1073/pnas.85.18.6612. PMID: 3045820; PMCID: PMC282027.
[3]. Seelmeier S, Schmidt H, et,al. Human immunodeficiency virus has an aspartic-type protease that can be inhibited by pepstatin A. Proc Natl Acad Sci U S A. 1988 Sep;85(18):6612-6. doi: 10.1073/pnas.85.18.6612. PMID: 3045820; PMCID: PMC282027.
[4]. Yoshida H, Okamoto K, et,al.Pepstatin A, an aspartic proteinase inhibitor, suppresses RANKL-induced osteoclast differentiation. J Biochem. 2006 Mar;139(3):583-90. doi: 10.1093/jb/mvj066. PMID: 16567424.
[5]. Okada M, Irie S, S et,al. Pepstatin A induces extracellular acidification distinct from aspartic protease inhibition in microglial cell lines. Glia. 2003 Aug;43(2):167-74. doi: 10.1002/glia.10237. PMID: 12838508.
[6]. Matarrese P, Nencioni L, et,al. Pepstatin A alters host cell autophagic machinery and leads to a decrease in influenza A virus production. J Cell Physiol. 2011 Dec;226(12):3368-77. doi: 10.1002/jcp.22696. PMID: 21344392.
| Cas No. | 26305-03-3 | SDF | |
| 别名 | 抑肽素; Pepstatin A | ||
| 化学名 | 3-hydroxy-4-[2-[[3-hydroxy-6-methyl-4-[[3-methyl-2-[[3-methyl-2-(3-methylbutanoylamino)butanoyl]amino]butanoyl]amino]heptanoyl]amino]propanoylamino]-6-methylheptanoic acid | ||
| Canonical SMILES | CC(C)CC(C(CC(=O)O)O)NC(=O)C(C)NC(=O)CC(C(CC(C)C)NC(=O)C(C(C)C)NC(=O)C(C(C)C)NC(=O)CC(C)C)O | ||
| 分子式 | C34H63N5O9 | 分子量 | 685.9 |
| 溶解度 | ≥ 34.3mg/mL in DMSO | 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 | 1.4579 mL | 7.2897 mL | 14.5794 mL |
| 5 mM | 0.2916 mL | 1.4579 mL | 2.9159 mL |
| 10 mM | 0.1458 mL | 0.729 mL | 1.4579 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。
Pepstatin A: polymerization of an oligopeptide
Micron 1994;25(2):189-217.8055247 10.1016/0968-4328(94)90042-6
Pepstatin A, a pentapeptide with the molecular weight of 686, is a naturally occurring inhibitor of aspartyl proteases secreted by Streptomyces species. Above a critical concentration of 0.1 mM at low ionic strength and neutral pH, it can polymerize into filaments which may extend over several micrometers. After negative staining, these filaments show a helical substructure with characteristic diameters ranging from 6 to 12 nm. Selected images at higher magnification suggest the filaments are composed of two intertwined 6 nm strands. This is in agreement with the optical diffraction analysis which additionally established a periodic pitch of 25 nm for the helical intertwining. Rotary shadowing of the Pepstatin A filaments clearly demonstrated the right-handedness of the helical twist. In physiological salt solution or at higher concentrations of Pepstatin A, a variety of higher order structures were observed, including ribbons, sheets and cylinders with both regular and twisted or irregular geometries. Pepstatin A can interact with intermediate filament subunit proteins. These proteins possess a long, alpha-helical rod domain that forms coiled-coil dimers, which through both hydrophobic and ionic interactions form tetramers which, in turn, in the presence of physiological salt concentrations, polymerize into the 10 nm intermediate filaments. In the absence of salt, Pepstatin A and intermediate filament proteins polymerize into long filaments with a rough surface and a diameter of 15-17 nm. This polymerization appears to be primarily driven by nonionic interactions between Pepstatin A and polymerization-competent forms of intermediate filament proteins, resulting in a composite filament. Polymerization-incompetent proteolytic fragments of vimentin, lacking portions of the head and/or tail domain, failed to copolymerize with Pepstatin A into long filaments under these conditions. These peptides, as well as bovine serum albumin, were found to stick to the surface of Pepstatin A filaments, ribbons and sheets. Independent evidence for direct association of Pepstatin A with intermediate filament subunit proteins was provided not only by electron microscopy but also by UV difference spectra. Pepstatin A loses its ability to inhibit the aspartyl protease of the human immunodeficiency virus type 1 following polymerization into the higher order structures described here. The amazing fact that Pepstatin A can spontaneously self-associate to form very large polymers seems to be a more rare event for such small peptides. The other examples of synthetic or naturally occurring oligopeptides discussed in this review which are able to polymerize into higher order structures possess a common property, their hydrophobicity, often manifested by clusters of valine or isoleucine residues.(ABSTRACT TRUNCATED AT 400 WORDS)
Aspartic protease-pepstatin A interactions: Structural insights on the thermal inactivation mechanism
Biochimie 2021 Oct;189:26-39.34116131 10.1016/j.biochi.2021.06.002
Aspartic proteases are the targets for structure-based drug design for their role in physiological processes and pharmaceutical applications. Structural insights into the thermal inactivation mechanism of an aspartic protease in presence and absence of bound Pepstatin A have been obtained by kinetics of thermal inactivation, CD, fluorescence spectroscopy and molecular dynamic simulations. The irreversible thermal inactivation of the aspartic protease comprised of loss of tertiary and secondary structures succeeded by the loss of activity, autolysis and aggregation The enthalpy and entropy of thermal inactivation of the enzyme in presence of Pepstatin A increased from 81.2 to 148.5 kcal mol-1, and from 179 to 359 kcal mol-1 K-1 respectively. Pepstatin A shifted the mid-point of thermal inactivation of the protease from 58 掳C to 77 掳C. The association constant (K) for Pepstatin A with aspartic protease was 2.5 卤 0.3 脳 10 5 M-1 and 螖Go value was -8.3 kcal mol-1. Molecular dynamic simulation studies were able to delineate the role of Pepstatin A in stabilizing backbone conformation and side chain interactions. In the C伪-backbone, the short helical segments and the conserved glycines were part of the most unstable segments of the protein. Understanding the mechanism of thermal inactivation has the potential to develop re-engineered thermostable proteases.
Inhibition of XMRV and HIV-1 proteases by Pepstatin A and acetyl-pepstatin
FEBS J 2012 Sep;279(17):3276-86.22804908 PMC6290463
The kinetic properties of two classical inhibitors of aspartic proteases (PRs), Pepstatin A and acetyl-pepstatin, were compared in their interactions with HIV-1 and xenotropic murine leukemia virus related virus (XMRV) PRs. Both compounds are substantially weaker inhibitors of XMRV PR than of HIV-1 PR. Previous kinetic and structural studies characterized HIV-1 PR-acetyl-pepstatin and XMRV PR-pepstatin A complexes and suggested dramatically different binding modes. Interaction energies were calculated for the possible binding modes and suggested a strong preference for the one-inhibitor binding mode for HIV-1 PR-acetyl-pepstatin and the two-inhibitor binding mode for XMRV PR-pepstatin A interactions. Comparison of the molecular models suggested that in the case of XMRV PR the relatively unfavorable interactions at S3' and the favorable interactions at S4 and S4' sites with the statine residues may shift the ground state binding towards the two-inhibitor binding mode, whereas the single molecule ground state binding of statines to the HIV-1 PR appear to be more favorable. The preferred single molecular binding to HIV-1 PR allows the formation of the transition state complex, represented by substantially better binding constants. Intriguingly, the crystal structure of the complex of acetyl-pepstatin with XMRV PR has shown a mixed type of binding: the unusual binding mode of two molecules of the inhibitor to the enzyme, in a mode very similar to the previously determined complex with Pepstatin A, together with the classical binding mode found for HIV-1 PR. The structure is thus in good agreement with the very similar interaction energies calculated for the two types of binding.
Pepstatin pull-down at high pH is a powerful tool for detection and analysis of napsin A
Biochem Biophys Res Commun 2019 Jul 12;515(1):145-148.31130231 10.1016/j.bbrc.2019.05.094
Napsin A is an intracellular aspartic protease and biomarker of various malignancies like lung adenocarcinoma and ovarian clear cell carcinoma, but its detection is usually limited to immunohistochemical techniques gaining excellent information on its distribution but missing information about posttranslational modifications (e.g. maturation state) of the protein. We present a protocol for specific enrichment of napsin A from clinical or biological specimens, that facilitates detailed analysis of the protein. By using the exceptionally broad pH range under which napsin A binds to its inhibitor Pepstatin A we achieve highly selective binding of napsin A while other aspartic proteases have negligible affinity. Using this method we demonstrate that lung napsin A in many mammals is a heterogeneous enzyme with a characteristic ladder-like appearance in SDS-PAGE that might be caused by proteolytically processed N- and/or C-termini, in contrast to the more homogeneous form found in kidneys and primary lung adenocarcinoma.
A multimodal Pepstatin A peptide-based nanoagent for the molecular imaging of P-glycoprotein in the brains of epilepsy rats
Biomaterials 2016 Jan;76:173-86.26524537 10.1016/j.biomaterials.2015.10.050
Regional overexpression of the multidrug transporter P-glycoprotein (P-gp) in epileptic brain tissues may lower antiepileptic drugs concentrations at the target site and contribute to pharmacoresistance in refractory epilepsy. However, few techniques are available to quantitate the level of P-gp expression noninvasively in vivo. In this study, we developed a nanoagent by conjugating superparamagnetic iron oxide nanoparticles with a near infrared probe and the targeting element Pepstatin A, a peptide with specific affinity for P-gp. In a rat model of epilepsy, the nanoagent was readily and selectively accumulated within epileptogenic cerebral regions, which were detectable by both magnetic resonance imaging and optical imaging modalities. This P-gp-targeted nanoagent could be used not only in the molecular imaging of P-gp expression changes in seizure-induced regional, understanding the mechanisms of P-gp disorders, and the prediction of refractory epilepsy, but also in targeted therapies with P-gp modulators.