Parasin I
(Synonyms: 鲶鱼抗菌肽 I) 目录号 : GC32348
ParasinI是一种由19个氨基酸残基组成的多肽，源于从鲶鱼皮肤分离到的组蛋白H2A，具有抗菌的作用。
Cas No.:219552-69-9
Sample solution is provided at 25 µL, 10mM.
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Quality Control & SDS
- View current batch:
- Purity: >98.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Cell experiment: | The antimicrobial activity of each peptide is determined using the broth microdilution assay. Briefly, single colonies of bacteria and fungi are inoculated into 3% trypticase soy broth (TSB) and Saboraud's medium, respectively, and cultured overnight at 37 and 30°C, respectively. Aliquots of each culture are transferred to 50 mL of fresh medium and incubated for an additional 3-6 h to obtain midlogarithmic phase cells. The cells are then washed and resuspended in 10 mM sodium phosphate buffer (NAPB), pH 7.4. The relationship between absorbance at 620 nm and colony-forming units (cfus) is determined for each microorganism by spreading serial dilutions of the cell suspension onto TSB or Saboraud agar plates. The cell suspension is diluted to 5×105 cfu/mL with 10 mM NAPB. Each well of 96- well propylene microtiter plates is filled with 90 mL of the diluted suspension and 10 mL of serially diluted peptide samples. After incubation for 3 h, fresh medium is added to the mixture and incubated at 37°C (bacteria) or 30°C (fungi) for an additional 16 h. The inhibition of growth is determined by measuring absorbance at 620 nm with a Model 550 Microplate Reader. The lowest concentration of peptide that completely inhibits growth is defined as the ‘minimal inhibitory concentration’ (MIC). The MICs are the average values obtained in triplicates in three independent experiments. |
References: [1]. Koo YS, et al. Structure-activity relations of parasin I, a histone H2A-derived antimicrobial peptide. Peptides. 2008 Jul;29(7):1102-8. | |
Parasin I is a 19-amino acid histone H2A-derived peptide isolated from the skin of the catfish, and shows antimicrobial activity.
Parasin I with comparable antimicrobial activities localized to the cell membrane and subsequently permeabilized the outer and cytoplasmic membranes. Parasin I and its active analogs show strong cytoplasmic membrane permeabilizing activity[1]. Codon optimized parasin I fused with human lysozyme is expressed in Pichia pastoris, and has potent antibiotic activity[2].
[1]. Koo YS, et al. Structure-activity relations of parasin I, a histone H2A-derived antimicrobial peptide. Peptides. 2008 Jul;29(7):1102-8. [2]. Zhao H, et al. Characterization of bioactive recombinant antimicrobial peptide parasin I fused with human lysozyme expressed in the yeast Pichia pastoris system. Enzyme Microb Technol. 2015 Sep;77:61-7.
| Cas No. | 219552-69-9 | SDF | |
| 别名 | 鲶鱼抗菌肽 I | ||
| Canonical SMILES | Lys-Gly-Arg-Gly-Lys-Gln-Gly-Gly-Lys-Val-Arg-Ala-Lys-Ala-Lys-Thr-Arg-Ser-Ser | ||
| 分子式 | C82H154N34O24 | 分子量 | 2000.31 |
| 溶解度 | Soluble 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 | 0.4999 mL | 2.4996 mL | 4.9992 mL |
| 5 mM | 0.1 mL | 0.4999 mL | 0.9998 mL |
| 10 mM | 0.05 mL | 0.25 mL | 0.4999 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。
Parasin I, an antimicrobial peptide derived from histone H2A in the catfish, Parasilurus asotus
FEBS Lett 1998 Oct 23;437(3):258-62.PMID:9824303DOI:10.1016/s0014-5793(98)01238-1.
In response to epidermal injury, Parasilurus asotus, a catfish, secreted a strong antimicrobial peptide into the epithelial mucosal layer. The molecular mass of the antimicrobial peptide, named Parasin I, was 2000.4 Da, as determined by matrix-associated laser desorption ionization mass spectrometry. The complete amino acid sequence of Parasin I, which was determined by automated Edman degradation, was Lys-Gly-Arg-Gly-Lys-Gln-Gly-Gly-Lys-Val-Arg-Ala-Lys-Ala-Lys-Thr-Arg-Ser- Ser. Eighteen of the 19 residues in Parasin I were identical to the N-terminal of buforin I, a 39-residue antimicrobial peptide derived from the N-terminal of toad histone H2A [Kim et al. (1996) Biochem. Biophys. Res. Commun. 229, 381-387], which implies that Parasin I was cleaved off from the N-terminal of catfish histone H2A. Parasin I showed strong antimicrobial activity, about 12-100 times more potent than magainin 2, against a wide spectrum of microorganisms, without any hemolytic activity. Circular dichroism spectra of Parasin I indicated a structural content of 11% alpha-helix, 33% beta-sheet, and 56% random coils. The beta-sheet axial projection diagram of Parasin I showed an amphipathic structure. Our results indicate that the catfish may produce Parasin I from its histone H2A by a specific protease upon injury to protect against invasion by microorganisms.
Structure-activity relations of Parasin I, a histone H2A-derived antimicrobial peptide
Peptides 2008 Jul;29(7):1102-8.PMID:18406495DOI:10.1016/j.peptides.2008.02.019.
The structure-activity relations and mechanism of action of Parasin I, a 19-amino acid histone H2A-derived antimicrobial peptide, were investigated. Parasin I formed an amphipathic alpha-helical structure (residues 9-17) flanked by two random coil regions (residues 1-8 and 18-19) in helix-promoting environments. Deletion of the lysine residue at the N-terminal [Pa(2-19)] resulted in loss of antimicrobial activity, but did not affect the alpha-helical content of the peptide. The antimicrobial activity was recovered when the lysine residue was substituted with another basic residue, arginine ([R(1)]Pa), but not with polar, neutral, or acidic residues. Progressive deletions from the C-terminal [Pa(1-17), Pa(1-15)] slightly increased the antimicrobial activity (1-4 microg/ml) without affecting the alpha-helical content of the peptide. However, further deletion [Pa(1-14)] resulted in nearly complete loss of antimicrobial activity and alpha-helical structure. Confocal microscopic analysis and membrane permeabilization assays showed that Parasin I and its analogs with comparable antimicrobial activities localized to the cell membrane and subsequently permeabilized the outer and cytoplasmic membranes. Pa(1-14) also localized to the cell membrane, but lost membrane-permeabilizing activity, whereas Pa(2-19) showed poor membrane-binding and -permeabilizing activities. The results indicate that the basic residue at the N-terminal is essential for the membrane-binding activity of Parasin I, and among the membrane-binding Parasin I analogs, the alpha-helical structure is necessary for the membrane-permeabilizing activity.
Cathepsin D produces antimicrobial peptide Parasin I from histone H2A in the skin mucosa of fish
FASEB J 2002 Mar;16(3):429-31.PMID:11821259DOI:10.1096/fj.01-0736fje.
Parasin I is a potent 19-residue antimicrobial peptide isolated from the skin mucus of wounded catfish (Parasilurus asotus). Here we describe the mechanism of Parasin I production from histone H2A in catfish skin mucosa on epidermal injury. Cathepsin D is found to exist in the mucus as an inactive proenzyme (procathepsin D), and a metalloprotease, induced on injury, cleaves procathepsin D to generate active cathepsin D. This activated form of cathepsin D then cleaves the Ser19-Arg20 bond of histone H2A to produce Parasin I. Immunohistochemical analysis reveals that unacetylated histone H2A, a precursor of Parasin I, and procathepsin D are present in the cytoplasm of epithelial mucous cells and that Parasin I is produced on the mucosal surface on epidermal injury. Western blot analysis shows that Parasin I is also present in the skin mucus of other fish species. Furthermore, Parasin I shows good antimicrobial activity against fish-specific bacterial pathogens. Taken together, these results indicate that cathepsin D and a metalloprotease participate in the production of Parasin I from histone H2A and that Parasin I contributes to the innate host defense of the fish against invading microorganisms.
Characterization of bioactive recombinant antimicrobial peptide Parasin I fused with human lysozyme expressed in the yeast Pichia pastoris system
Enzyme Microb Technol 2015 Sep;77:61-7.PMID:26138401DOI:10.1016/j.enzmictec.2015.06.001.
Parasin I (PI) is a 19 amino acid peptide with potent antimicrobial activities against a broad spectrum of microorganisms and is a good candidate for development as a novel antimicrobial agent. The objective of this study was to express and characterize a codon optimized Parasin I peptide fused with human lysozyme (hLY). A 513 bp cDNA fragment encoding the mature hLY protein and Parasin I peptide was designed and synthesized according to the codon bias of Pichia pastoris. A 4脳Gly flexible amino acid linker with an enterokinase cleavage (DDDDK) was designed to link the PI to the C-terminal of hLY. The codon optimized recombinant hLY-PI was cloned into the pPICZ伪A vector and expressed in P. pastoris. The over-expressed extracellular rehLY-PI was purified using Ni sepharose affinity column and exhibited a molecular mass of approximately 18 kDa. After digested with enterokinase the rehLY-PI protein release its corresponding rehLY and rePI, with molecular mass of 16 kDa and 2 kDa, respectively, on Tricine-SDS-PAGE. The released rehLY exhibited similar lytical activity against Micrococcus lysodeikticus to its commercial hLY. The digested rehLY-PI product exhibited antimicrobial activities against Bacillus subtilis, Staphylococcus aureus and Escherichia coli, and synergism has been found between the released rePI and rehLY. In conclusion, we successfully optimized a rehLY-PI fusion protein encoding gene and over-expressed the rehLY-PI in P. pastoris. The recombination protein digested with enterokinase released functional hLY and antimicrobial Parasin I, which demonstrates a potential for future use as an animal feed additive to partly replace antibiotic.
Nanoparticle Emulsions Enhance the Inhibition of NLRP3
Int J Mol Sci 2022 Sep 5;23(17):10168.PMID:36077562DOI:10.3390/ijms231710168.
Antibacterial delivery emulsions are potential materials for treating bacterial infections. Few studies have focused on the role and mechanism of emulsions in inflammation relief. Therefore, based on our previous analysis, in which the novel and natural Pickering emulsions stabilized by antimicrobial peptide nanoparticles were prepared, the regulation effect of emulsion on inflammasome was explored in silico, in vitro and in vivo. Firstly, the interactions between inflammasome components and Parasin I or Pickering emulsion were predicted by molecular docking. Then, the inflammasome stimulation by different doses of the emulsion was tested in RAW 264.7 and THP-1 cells. Finally, in Kunming mice with peritonitis, NLRP3 and IL-1尾 expression in the peritoneum were evaluated. The results showed that the Pickering emulsion could combine with ALK, casp-1, NEK7, or NLRP3 to affect the assembly of the NLRP3 and further relieve inflammation. LPNE showed a dose-dependent inhibition effect on the release of IL-1尾 and casp-1. With the concentration of Parasin I increased from 1.5 mg/mL to 3 mg/mL, the LDH activity decreased in the chitosan peptide-embedded nanoparticles emulsion (CPENE) and lipid/peptide nanoparticles emulsion (LPNE) groups. However, from 1.5 to 6 mg/mL, LPNE had a dose-dependent effect on the release of casp-1. The CPENE and parasin I-conjugated chitosan nanoparticles emulsion (PCNE) may decrease the release of potassium and chloride ions. Therefore, it can be concluded that the LPNE may inhibit the activation of the inflammasome by decreasing LDH activity, potassium and chloride ions through binding with compositions of NLRP3.