53-79-2

Usage

Uses

1. Used in Biomedical Research:
PUROMYCIN is used as a research tool for studying protein synthesis, as it inhibits protein synthesis by binding to RNA and causing premature chain termination during translation in the ribosomes.
2. Used in Genetic Engineering:
PUROMYCIN is used as a selection agent for cells modified by genetic engineering, particularly for the selection of cell types harboring plasmids carrying puromycin resistance genes.
3. Used in Antineoplastic and Antitrypanosomal Applications:
PUROMYCIN is used as an antineoplastic and antitrypanosomal agent, exhibiting activity against various types of cancer and trypanosomal infections.
4. Used in Cell Biology:
PUROMYCIN is used as a tool in cell biology to select mammalian cell lines that have been transformed by vectors expressing puromycin-N-acetyl-transferase.
5. Used in Biochemistry and Nephrology Research:
PUROMYCIN is used as a reversible inhibitor of dipeptidyl-peptidase II (serine peptidase) and cytosol alanyl aminopeptidase (metallopeptidase), making it extremely useful in distinguishing between aminopeptidase M (active) and cytosol alanyl aminopeptidase (inhibited by puromycin).
6. Used in mRNA Display:
PUROMYCIN is used in mRNA display, allowing the growing peptide chain to be covalently bonded to its own mRNA template, which is critical for the technique's success.

Hazard

Toxic to living cells of all kinds.

Toxicity evaluation

Puromycin is a specific metabolic inhibitor of protein synthesis and acts as an aminoacyl-tRNA analog and peptidyl acceptor. The latter causes premature chain termination of the protein and the release of nascent or growing polypeptide chains. In liver, it has been shown to cause fat accumulation without causing death of the hepatocytes. Puromycin causes focal glomerular sclerosis and alters the morphology, localization of anionic sites, and metabolism of renal epithelial cells. This injury is attributable to the production of reactive oxygen species.

InChI:InChI=1/C22H29N7O5.2ClH/c1-28(2)19-17-20(25-10-24-19)29(11-26-17)22-18(31)16(15(9-30)34-22)27-21(32)14(23)8-12-4-6-13(33-3)7-5-12;;/h4-7,10-11,14-16,18,22,30-31H,8-9,23H2,1-3H3,(H,27,32);2*1H/t14-,15+,16+,18+,22?;;/m0../s1

Upstream product

• 57182-86-2 6-dimethylamino-9-[3'-(O-methyl) (2S)-N-benzyloxycarbonyltyrosinylamino-3'-deoxy-β-D-ribofuranosyl]purine 
• 58-58-2 puromycin dihydrogen chloride 
• 111210-69-6 O-methyl-N-phthaloyl-(S)-tyrosyl chloride 
• 60-18-4 L-tyrosine 
• 17554-34-6 (2S)-2-[(benzyloxycarbonyl)amino]-3-(4-methoxyphenyl)propanoic acid 
• 1164-16-5 Z-L-Tyr-OH 
• 58-60-6 3'-amino-3'-deoxy-N⁶,N⁶-dimethyladenosine 
• 384334-58-1 2,2,2-Trifluoro-N-[(2S,3S,4R,5R)-4-hydroxy-2-hydroxymethyl-5-(6-[1,2,4]triazol-4-yl-purin-9-yl)-tetrahydro-furan-3-yl]-acetamide 
• 749200-34-8 9-(3-azido-3-deoxy-β-D-ribofuranosyl)-6-(1,2,4-triazol-4-yl)purine 
• 384334-64-9 9-(3-azido-3-deoxy-β-D-ribofuranosyl)-6-(dimethylamino)purine 
• 2504-55-4 3'-amino-3'-deoxyadenosine 
• 2627-64-7 9-(2,3-anhydro-β-D-ribofuranosyl)adenine 
• 68044-56-4 3'-Trifluoracetamido-3'-desoxyadenosin 
• 57183-05-8 tert-butyl [(S)-1-({(2S,3S,4R,5R)-5-[6-(dimethylamino)-9H-purin-9-yl]-4-hydroxy-2-(hydroxymethyl)tetrahydrofuran-3-yl}amino)-3-(4-methoxyphenyl)-1-oxopropan-2yl]carbamate 

Downstream Products

• 57183-05-8 tert-butyl [(S)-1-({(2S,3S,4R,5R)-5-[6-(dimethylamino)-9H-purin-9-yl]-4-hydroxy-2-(hydroxymethyl)tetrahydrofuran-3-yl}amino)-3-(4-methoxyphenyl)-1-oxopropan-2yl]carbamate 
• 151967-82-7 [(S)-1-[(2S,3S,4R,5R)-5-(6-Dimethylamino-purin-9-yl)-4-hydroxy-2-hydroxymethyl-tetrahydro-furan-3-ylcarbamoyl]-2-(4-methoxy-phenyl)-ethyl]-carbamic acid 9H-fluoren-9-ylmethyl ester 
• 1477500-67-6 C₄₈H₅₈N₉O₉P 
• 1477500-65-4 C₃₉H₄₁N₇O₈ 
• 1477500-63-2 C₆₀H₅₉N₇O₁₀ 
• 151967-83-8 [(S)-1-[(2S,3S,4R,5R)-2-[Bis-(4-methoxy-phenyl)-phenyl-methoxymethyl]-5-(6-dimethylamino-purin-9-yl)-4-hydroxy-tetrahydro-furan-3-ylcarbamoyl]-2-(4-methoxy-phenyl)-ethyl]-carbamic acid 9H-fluoren-9-ylmethyl ester 

Relevant academic research and scientific papers

Radiosynthesis of Carbon-11 Labeled Puromycin as a Potential PET Candidate for Imaging Protein Synthesis in Vivo

In order to address the limitations associated with the present range of PET radiotracers used for imaging protein synthesis in vivo we have synthesized a candidate PET radiotracer based on Puromycin (3, PURO), a protein synthesis inhibitor. The desmethylPURO 9 precursor for radiolabeling with carbon-11 radioisotope was synthesized in two steps employing EDC/HOBt amide coupling in overall 76% yield. Optimal conditions for radiolabeling were then established via methylation/deprotection sequence. Under these conditions as determined by NMR analysis 9 showed partial stability (ca. 80%) under acidic conditions. Limited evidence of stereochemical stability of 3 was also found. The radiolabeling of intermediate [11C]12 was accomplished with up to 57% conversion from [11C]iodomethane. An automated method was then developed for high radioactivity radiosynthesis to produce [11C]3 ([11C]PURO) in 16 ± 6% (n = 3) decay corrected radiochemical yields.

Versatile Histochemical Approach to Detection of Hydrogen Peroxide in Cells and Tissues Based on Puromycin Staining

Hydrogen peroxide (H2O2) is a central reactive oxygen species (ROS) that contributes to diseases from obesity to cancer to neurodegeneration but is also emerging as an important signaling molecule. We now report a versatile histochemical approach for detection of H2O2 that can be employed across a broad range of cell and tissue specimens in both healthy and disease states. We have developed a first-generation H2O2-responsive analogue named Peroxymycin-1, which is based on the classic cell-staining molecule puromycin and enables covalent staining of biological samples and retains its signal after fixation. H2O2-mediated boronate cleavage uncages the puromycin aminonucleoside, which leaves a permanent and dose-dependent mark on treated biological specimens that can be detected with high sensitivity and precision through a standard immunofluorescence assay. Peroxymycin-1 is selective and sensitive enough to image both exogenous and endogenous changes in cellular H2O2 levels and can be exploited to profile resting H2O2 levels across a panel of cell lines to distinguish metastatic, invasive cancer cells from less invasive cancer and nontumorigenic counterparts, based on correlations with ROS status. Moreover, we establish that Peroxymycin-1 is an effective histochemical probe for in vivo H2O2 analysis, as shown through identification of aberrant elevations in H2O2 levels in liver tissues in a murine model of nonalcoholic fatty liver disease, thus demonstrating the potential of this approach for studying disease states and progression associated with H2O2. This work provides design principles that should enable development of a broader range of histochemical probes for biological use that operate via activity-based sensing.

Design of photocaged puromycin for nascent polypeptide release and spatiotemporal monitoring of translation

The antibiotic puromycin, which inhibits protein translation, is used in a broad range of biochemical applications. The synthesis, characterization, and biological applications of NVOC-puromycin, a photocaged derivative that is activated by UV illumination, are presented. The caged compound had no effect either on prokaryotic or eukaryotic translation or on the viability of HEK 293 cells. Furthermore, no significant release of ribosome-bound polypeptide chains was detected invitro. Upon illumination, cytotoxic activity, invitro translation inhibition, and polypeptide release triggered by the uncaging of NVOC-puromycin were equivalent to those of the commercial compound. The quantum yield of photolysis was determined to be 1.1±0.2 % and the NVOC-puromycin was applied to the detection of newly translated proteins with remarkable spatiotemporal resolution by using two-photon laser excitation, puromycin immunohistochemistry, and imaging in rat hippocampal neurons. On like a light: The antibiotic puromycin (green) is a translation inhibitor that triggers the release of the nascent polypeptide chain (red) from the ribosome (yellow) and it is used in a number of applications. A photocaged puromycin derivative, NVOC-puromycin, was synthesized and characterized. Both functional recovery upon UV illumination and biological inactivity invitro and invivo were demonstrated.

ESTERASES FOR MONITORING PROTEIN BIOSYNTHESIS IN VITRO

The present invention relates to the use of an esterase for monitoring and/or tracking the synthesis of a protein, polypeptide or peptide in a cell-free translation system or in an in vivo expression system in which the synthesis of a protein, polypeptide or peptide can occur, wherein said monitoring and/or tracking comprises the detection of the function of said esterase. The present invention further relates to a vector comprising a nucleic acid molecule coding for an esterase and expressing an esterase fusionprotein. Moreover, the present invention relates to a vector comprising a nucleic acid molecule coding for an esterase and comprising in frame at least one multiple cloning site for a further protein/polypeptide/peptide, to be expressed in form of a fusion protein comprising said esterase (esterase activity) and said further proteinaceous peptide structure. The present invention also provides for a protein, polypeptide or peptide encoded by the vectors of the present invention. Additionally, the present invention relates to a kit comprising a vector of the present invention or a nucleic acid molecule as comprised by the vectors of the present invention. Also disclosed is a method for monitoring and/or tracking the synthesis of a protein, polypeptide or peptide in a cell-free translation system or in an in vivo expression system, comprising the step of detecting the function of an esterase. The present invention also teaches a method for immobilising a protein, polypeptide or peptide comprising the steps of (a) tagging said protein, polypeptide or peptide with an esterase and (b) binding said esterase to an esterase inhibitor, wherein said esterase inhibitor is immobilized on a solid substrate. Moreover, the present invention relates to uses of the vectors of the present invention or the nucleic acid molecules comprised therein for the preparation of a kit or for monitoring and/or tracking the synthesis of a protein, polypeptide or peptide in a cell-free translation system, whereby the monitoring and/or tracking comprises the detection of the function of said esterase.

Syntheses of puromycin from adenosine and 7-deazapuromycin from tubercidin, and biological comparisons of the 7-aza/deaza pair

Protection (05′) of 2′,3′-anhydroadenosine with tert-butyldiphenylsilyl chloride and epoxide opening with dimethylboron bromide gave the 3′-bromo-3′-deoxy xylo isomer which was treated with benzylisocyanate to give the 2′-O-(N-benzylcarbamoyl) derivative. Ring closure gave the oxazolidinone, and successive deprotection concluded an efficient route to 3′-amino-3′-deoxyadenosine. Analogous treatment ofthe antibiotic tubercidin {7-deazaadenosine; 4-amino-7-(β-D-ribofuranosyl)-pyrrolo[2,3-d]pyrimidine} gave 3′-amino-3′-deoxytubercidin. Trifluoroacetylation of the 3′-amino function, elaboration of the heterocyclic amino group into a (1,2,4-triazol-4-yl) ring with N,N′-bis-[(dimethylamino)methylene]hydrazine, and nucleophilic aromatic substitution with dimethylamine gave puromycin aminonucleoside [9-(3-amino-3-deoxy-β-D-ribofuranosyl)-6-(dimethylamino)purine] and its 7-deaza analogue. Aminoacylation [BOC-(4-methoxy-L-phenylalanine)] and deprotection gave puromycin and 7-deazapuromycin. Most reactions gave high yields at or below ambient temperature. Equivalent inhibition of protein biosynthesis in a rabbit reticulocyte system and parallel growth inhibition of several bacteria were observed with the 7-aza/deaza pair. Replacement of N7 in the purine ring of puromycin by "CH" has no apparent effect on biological activity.

Synthesis of 6-Dimethylamino-9-[3'-(O-Methyl) (2S)-[UL-14C]Tyrosinylamino)-3'-Deoxy-β-D- ribofuranosyl] purine

In order to investigate and further refine the mechanism of the unique cleavage activity of the 18 amino acid 2A region of the foot-and-mouth-disease virus (FMDV), the synthesis of 14C-labelled puromycin is required. Puromycin is an inhibitor of protein synthesis and is an analogue of the terminal aminoacyl-adenosine portion of aminoacyl-tRNA. A short and expedient four step synthesis of 6-dimethylamino-9-[3'-(O-methyl) (2S)-[UL-14C]tyrosinylamino)-3'-deoxy-β-D-ribofuranos purine (14C-labelled puromycin) starting from (2S)-[UL-14C]-tyrosine is therefore described.