DiZPK Hydrochloride
目录号 : GC19125
DiZPK Hydrochloride是一种光反应性赖氨酸类似物，含有光激活的二吖丙啶官能团，能够作为光交联剂促进原核和真核细胞中蛋白质-蛋白质相互作用。
Cas No.:2349295-23-2
Sample solution is provided at 25 µL, 10mM.
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DiZPK Hydrochloride is a photoreactive lysine analog containing a photoactivated diaziridine functional group that can promote covalent photocrosslinking of protein-protein interactions in prokaryotic and eukaryotic cells[1].
DiZPK Hydrochloride has two characteristics as a probe: first, as a derivative of pyrrolysine, DiZPK Hydrochloride can be used in genetic codon expansion technology; second, by modifying the pyrrolysine-TrNA synthase (PylRS) and its corresponding tRNA (tRNAPyl) system, non-natural amino acids can be introduced into the stop codon TAG position. In addition, DiZPK Hydrochloride contains a diacridine photocrosslinking group. Under specific 365nm wavelength irradiation, diacridine can be converted into free radical groups, thereby achieving transient protein-protein non-covalent conversion to covalent interaction. Compared with other photocrosslinking groups, the "short" structure of the diacridine linker can minimize the impact on the function of the target protein[2].
DiZPK Hydrochloride achieved a cross-linking efficiency of 60% within 1 minute of UV irradiation and a maximum efficiency of over 80% within 3 minutes, indicating that DiZPK has high flexibility[3].
By introducing the DiZPK photoaffinity probe site-specifically into HdeA, it was combined with gel-based proteomics to enable substrate analysis of HdeA in living Escherichia coli cells at very low pH. Two important periplasmic partners, DegP and SurA, were found among the natural substrates identified by mass spectrometry, which were initially protected by HdeA at low pH but then assisted the refolding of other proteins mediated by HDEa after neutralization[3].
References:
[1]. Chengjie Chen, Tao Peng.Protocol for Site-Specific Photo-Crosslinking Proteomics to Identify Protein-Protein Interactions in Mammalian Cells.STAR Protoc.2020 Sep15;1(3):100109.doi: 10.1016/j.xpro.2020.100109. eCollection 2020 Dec 18.
[2]. He D, Xie X, et,al. Quantitative and Comparative Profiling of Protease Substrates through a Genetically Encoded Multifunctional Photocrosslinker. Angew Chem Int Ed Engl. 2017 Nov 13;56(46):14521-14525. doi: 10.1002/anie.201708151. Epub 2017 Oct 11. PMID: 28940571.
[3]. Zhang M, Lin S, et,al. A genetically incorporated crosslinker reveals chaperone cooperation in acid resistance. Nat Chem Biol. 2011 Sep 4;7(10):671-7. doi: 10.1038/nchembio.644. PMID: 21892184.
DiZPK Hydrochloride是一种光反应性赖氨酸类似物，含有光激活的二吖丙啶官能团，能够促进原核和真核细胞中蛋白质-蛋白质相互作用的共价光交联[1]。
DiZPK Hydrochloride作为探针具有两个特点：其一，DiZPK Hydrochloride作为吡咯赖氨酸的衍生物，可用于遗传密码子扩展技术；其二，通过对吡咯赖氨酸-TrNA合酶（PylRS）及其对应的tRNA（tRNAPyl）系统的改造，可将非天然氨基酸引入终止密码子TAG位置。此外，DiZPK Hydrochloride含有双吖啶光交联基团，在特定的365nm波长照射下，双吖啶可转化为自由基基团，从而实现瞬时蛋白-蛋白非共价转化为共价作用。与其他光交联基团相比，双吖啶Linker的“短”结构可最大程度地减少对靶蛋白功能的影响[2]。
DiZPK Hydrochloride在紫外线照射1分钟内交联效率达到60%，3分钟内达到最高效率 80%以上，表明DiZPK具有很高的灵活性[3]。
通过将DiZPK光亲和探针位点特异性引入HdeA，它与基于凝胶的蛋白质组学相结合，可以在极低pH值下对活大肠杆菌细胞中的HdeA进行底物分析。在质谱鉴定的天然底物中发现了两个重要的周质伙伴DegP和SurA，它们最初在低pH值下受到HdeA的保护，但随后在中和后协助HDEa介导的其他蛋白质的重新折叠[3]。
| Assessing the crosslinking efficiency of DiZPK Hydrochloride [1]: | |
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Preparation Method |
Assessing the crosslinking efficiency of DiZPK Hydrochloride by analyzing the amounts of crosslinked products produced after they were introduced at a common position, Phe28, which is located at the dimer interface of the HdeA structure. The crosslinking was performed at neutral pH by exposing the E. coli cells, which heterologously expressed HdeA-His, and carried either of the two probes, to UV light (365 nm) for different time periods.The crosslinked dimer products of HdeA were then analyzed by SDS-PAGE and immunoblotting with a His-tag specific antibody. |
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Applications |
For DiZPK Hydrochloride revealed a crosslinking efficiency of 60% within 1 min of UV exposure and achieved a maximum efficiency higher than 80% within 3 min demonstrating the higher flexibility of DiZPK. |
| DiZPK probe to capture periplasmic proteins interacting with HdeA in E. coli cells [1]: | |
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Cell lines |
E. coli cells |
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Preparation Method |
Cells expressing HdeA proteins and carrying a singlesite incorporated DiZPK Hydrochloride at a variety of specific positions were grown at 37°C treated at pH 2.3 for 30 min before being subjected to 365nm light irradiation for 3 min. Immunoblotting analysis of lysates about the acid-treated cells. |
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Applications |
DiZPK Hydrochloride was introduced into the two hydrophobic regions of HdeA, there were multiple protein bands containing HdeA. These bands showed higher molecular weight than the HdeA monomer and thus represented the cross-linked form of HdeA. |
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References: [1]. Zhang M, Lin S, et,al. A genetically incorporated crosslinker reveals chaperone cooperation in acid resistance. Nat Chem Biol. 2011 Sep 4;7(10):671-7. doi: 10.1038/nchembio.644. PMID: 21892184. |
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| Cas No. | 2349295-23-2 | SDF | |
| Canonical SMILES | N[C@H](C(O)=O)CCCCNC(NCCCC1(N=N1)C)=O.Cl | ||
| 分子式 | C12H24ClN5O3 | 分子量 | 321.8 |
| 溶解度 | Water : 8.33 mg/mL (25.89 mM);DMSO : < 1 mg/mL (insoluble or slightly soluble) | 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 | 3.1075 mL | 15.5376 mL | 31.0752 mL |
| 5 mM | 0.6215 mL | 3.1075 mL | 6.215 mL |
| 10 mM | 0.3108 mL | 1.5538 mL | 3.1075 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