Bottromycin A2
目录号 : GC48365
An antibiotic
Cas No.:15005-62-6
Sample solution is provided at 25 µL, 10mM.
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Quality Control & SDS
- View current batch:
- Purity: >80.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Bottromycin A2 is an antibiotic originally isolated from Streptomyces.1 It blocks the binding of aminoacyl-tRNA to the A site of the 50S ribosome, inhibiting protein synthesis.2 Bottromycin A2 is active against clinical isolates of methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococci with MIC values of 1 and 0.5 µg/ml, respectively.
1.Nakamura, S., Yajima, T., Lin, Y., et al.Isolation and characterization of bottromycins A2, B2, C2J. Antibiot. (Tokyo)20(1)1-5(1967) 2.Gouda, H., Kobayashi, Y., Yamada, T., et al.Three-dimensional solution structure of bottromycin A2: A potent antibiotic active against methicillin-resistant Staphylococcus aureus and vancomycin-resistant EnterococciChem. Pharm. Bull. (Tokyo)60(2)169-171(2012)
| Cas No. | 15005-62-6 | SDF | |
| Canonical SMILES | O=C1[C@@]2([H])N(CC[C@H]2C)C(C/N=C([C@@H](NC([C@@H](N1)C(C)C)=O)C(C)(C)C)/N[C@@H](C(C)(C)C)C(N[C@H](C(N[C@@H](C3=NC=CS3)CC(OC)=O)=O)[C@H](C4=CC=CC=C4)C)=O)=O | ||
| 分子式 | C42H62N8O7S | 分子量 | 823.1 |
| 溶解度 | Ether: soluble,Methanol: soluble,Water: soluble | 储存条件 | -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 | 1.2149 mL | 6.0746 mL | 12.1492 mL |
| 5 mM | 0.243 mL | 1.2149 mL | 2.4298 mL |
| 10 mM | 0.1215 mL | 0.6075 mL | 1.2149 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。
Mode of action of Bottromycin A2: effect of Bottromycin A2 on polysomes
FEBS Lett 1983 Mar 7;153(1):53-9.PMID:6337880DOI:10.1016/0014-5793(83)80118-5.
When Bottromycin A2 was added to an in vitro protein synthesis system carried out by naturally occurring polysomes, it inhibited protein synthesis effectively. Examination of the 3 steps of peptide chain elongation revealed that the binding of aminoacyl-tRNA to the polyribosomes was inhibited by Bottromycin A2. In contrast, we concluded that the peptide bond formation and the translocation steps in this system were not inhibited by Bottromycin A2 on the basis of the following observations: (1) The break-down of polysomes, which is dependent on EFG, puromycin and RR (ribosome releasing) factor, was insensitive to Bottromycin A2; (2) The puromycin dependent release of polypeptide from polysomes, with or without EFG, was not inhibited by Bottromycin A2. Thus bottromycin specifically interferes with proper functioning of the A sites of polysomes. This is consistent with the results obtained using the model system with synthetic polynucleotides.
Mode of action of Bottromycin A2. Release of aminoacyl- or peptidyl-tRNA from ribosomes
J Biol Chem 1976 Apr 25;251(8):2299-306.PMID:770464doi
Bottromycin A2 inhibited MS2 phage RNA-dependent protein synthesis as well as polyuridylic acid-(poly(U))- or polyadenylic acid (poly(A))-dependent polypeptide synthesis. When the ribosomal complex with N-acetyl-[14C]phenylalanyl-tRNA (N-acetyl-[14C]Phe-tRNA) at the A site was subjected to Bottromycin A2, the release of N-acetyl-[14C]Phe-tRNA was observed while no release of N-acetyl-[14C]Phe-tRNA from the donor site was observed, indicating that the action of Bottromycin A2 is specific to the A site of ribosomes. Due to bottromycin's capacity to release [14C]Phe-tRNA or N-acetyl-[14C]Phe-tRNA from the ribosomal acceptor site (A site), Bottromycin A2 inhibited the nonenzymatic binding of N-acetyl-[14C]Phe-tRNA and elongation factor T (EF-T)-dependent binding if the concentration of EF-Tu-GTP-[14C]Phe-tRNA ternary complex was low. Our data are consistent with the possibility that the inhibition of overall polypeptide synthesis by Bottromycin A2 is at least partly due to Bottromycin A2's activity to release aminoacyl- or oligopeptidyl-tRNA from ribosomes. Among 10 antibiotics tested, Bottromycin A2 and lincomycin released aminoacyl-tRNA from ribosomes.
Dissecting Bottromycin Biosynthesis Using Comparative Untargeted Metabolomics
Angew Chem Int Ed Engl 2016 Aug 8;55(33):9639-43.PMID:27374993DOI:10.1002/anie.201604304.
Bottromycin A2 is a structurally unique ribosomally synthesized and post-translationally modified peptide (RiPP) that possesses potent antibacterial activity towards multidrug-resistant bacteria. The structural novelty of bottromycin stems from its unprecedented macrocyclic amidine and rare β-methylated amino acid residues. The N-terminus of a precursor peptide (BtmD) is converted into Bottromycin A2 by tailoring enzymes encoded in the btm gene cluster. However, little was known about key transformations in this pathway, including the unprecedented macrocyclization. To understand the pathway in detail, an untargeted metabolomic approach that harnesses mass spectral networking was used to assess the metabolomes of a series of pathway mutants. This analysis has yielded key information on the function of a variety of previously uncharacterized biosynthetic enzymes, including a YcaO domain protein and a partner protein that together catalyze the macrocyclization.
Three-dimensional solution structure of Bottromycin A2: a potent antibiotic active against methicillin-resistant Staphylococcus aureus and vancomycin-resistant Enterococci
Chem Pharm Bull (Tokyo) 2012;60(2):169-71.PMID:22293474DOI:10.1248/cpb.60.169.
The three-dimensional (3D) structure of bottromycin A(2), a natural anti-methicillin-resistant Staphylococcus aureus (MRSA) and anti-vancomycin-resistant Enterococci (VRE) agent consisting of seven amino acids, has been investigated through NMR spectroscopy. On the basis of 57 experimental constraints, a total of 34 converged structures were obtained. The average pairwise atomic root mean square difference is 0.74±0.59 Å for all heavy atoms. The resulting structure indicates an interesting feature in that the three C-terminal residues of bottromycin A(2) fold back on the 12-membered cyclic skeleton made by the four N-terminal residues. Thus, MePro(2) and Thia-β-Ala-OMe(7), modification of which significantly affects the antibacterial activities of bottromycin A(2), are located on one side of its 3D structure. These distinct structural features might be important for the binding of bottromycin A(2) with the bacterial ribosome.
Thiazoline-Specific Amidohydrolase PurAH Is the Gatekeeper of Bottromycin Biosynthesis
J Am Chem Soc 2019 Jun 26;141(25):9748-9752.PMID:31192589DOI:10.1021/jacs.8b12231.
The ribosomally synthesized and post-translationally modified peptide (RiPP) Bottromycin A2 possesses potent antimicrobial activity. Its biosynthesis involves the enzymatic formation of a macroamidine, a process previously suggested to require the concerted efforts of a YcaO enzyme (PurCD) and an amidohydrolase (PurAH) in vivo. In vitro, PurCD alone is sufficient to catalyze formation of the macroamidine, but the process is reversible. We set out to probe the role of PurAH in macroamidine formation in vitro. We demonstrate that PurAH is highly selective for macroamidine-containing precursor peptides and cleaves C-terminal of a thiazoline, thus removing the follower peptide. After follower cleavage, macroamidine formation is irreversible, indicating PurAH as the gatekeeper of bottromycin biosynthesis. The structure of PurAH suggests residues involved in catalysis, which were probed through mutagenesis.