1. Antibiotic Puromycin ----An inhibitor of protein synthesis Ping Jiang 03-05-2000

2. Outline Discovery and Properties Structure and Functions Molecular Mechanism of Action Analogues and Synthesis Applications and Prospect

3. In 1952, the new antibiotic, Puromycin, isolated from the mold Streptomyces alboniger, has been found to be active against certain bacteria and Trypanosomes in Lederle laboratories division, American Cyanamid Company during the course of their antibiotic- screening program. One year later, the structure of puromycin was determined. Discovery Antibiotics and Chemotherapy, 1952, 2, 409 J. Am. Chem. Soc., 1953, 75, 2025

4. Chemical Properties Formula: C 22 H 29 N 7 O 5, 2HCl; MW: 544.2 pKa: 6.8 and 7.2 http://www.cayla.com/support/datasheets/protect.htm Puromycin is suitable for research purposes only. It must not be used on humans.

5. Structure Puromycin: a structural and a functional analogue of 3’ terminus of aminoacyl-tRNA The Ribosome: Structure, Function, and Evolution. 1990

6. Functions Many antibiotics target the ribosome. Puromycin tricks the ribosome into incorporating itself into the growing polypeptide chain causing premature termination, acting as an inhibitor of peptidyl transferase. The antibiotic inhibits the growth of Gram positive bacteria and various animal and insect cells. Fungi and Gram negative are resistant due to the low permeability of the antibiotic Problem: it kills eukaryotics and prokaryotics alike RNA, 2000, 6, 744-754; Inhibitors of Protein Synthesis, 1979

7. Binding Transpeptidation Translocation Mechanism:

8. puro A site P site Nascent Protein Release 5’ mRNA Stop coden Ribosome tRNA Nature,1963, 197, 1067-1977; FEBS Lett., 1997, 406, 223-233

9. Mechanism: The Molecular Basis of Antibiotic Action, 1972, 412

10. Analogues and Synthesis: A rich array of puromycin analogues with modified amino acid, carbohydrate, and/or base moieties has been prepared and evaluated biologically to study the structure-activity correlations….. Puromycin (1) and the N-acetylglucosaminyl asparagine-substitued puromycin analogue 2a-c

11. Analogues and Synthesis: Bioorg. & Medicinal Chem., 1995, 3, 1631-1636

12. Analogues and Synthesis: J. Org. Chem., 2001, 66, 8204-8210

13. Applications and Prospect: Puromycin has been widely used as a basic tool for studying protein synthesis. Now, puromycin hydrochloride is particularly useful for the selection of cell types harbouring plasmids carrying puromycin resistance genes. The specific bonding of puromycin to full-length protein at the C-terminus at a low concentration is potentially useful to the analysis of various biological phenomena. Nucleic Acid Research, 2000, 28, 1176-1182

14. Application and Prospect: puro A site P site Release 5’ mRNA Stop coden Ribosome tRNA Full length Protein Nucleic Acid Research, 2000, 28, 1176-1182