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Antibiotic Puromycin
----An inhibitor of protein synthesis
Ping Jiang
03-05-2000
Outline
• Discovery and Properties
• Structure and Functions
• Molecular Mechanism of Action
• Analogues and Synthesis
• Applications and Prospect
Discovery
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 antibioticscreening program. One year later, the structure of
puromycin was determined.
Antibiotics and Chemotherapy, 1952, 2, 409
J. Am. Chem. Soc., 1953, 75, 2025
Chemical Properties
Formula: C22H29N7O5, 2HCl;
MW:
544.2
pKa:
6.8 and 7.2
Puromycin is suitable for research purposes only.
It must not be used on humans.
http://www.cayla.com/support/datasheets/protect.htm
Structure
Puromycin: a structural and a functional analogue of
3’ terminus of aminoacyl-tRNA
The Ribosome: Structure, Function, and Evolution. 1990
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
Mechanism:
Binding
Translocation
Transpeptidation
Mechanism:
Nascent
Release
Protein
A site
P site
tRNA
puro
5’
Stop
coden
mRNA
Ribosome
Nature,1963, 197, 1067-1977;
FEBS Lett., 1997, 406, 223-233
Mechanism:
The Molecular Basis of Antibiotic Action, 1972, 412
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
Analogues and Synthesis:
Bioorg. & Medicinal Chem., 1995, 3, 1631-1636
Analogues and Synthesis:
J. Org. Chem., 2001, 66, 8204-8210
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
Application and Prospect:
Full length
Protein
Release
P site
tRNA
A site
puro
5’
mRNA
Stop
coden
Ribosome
Nucleic Acid Research, 2000, 28, 1176-1182