Cycloheximide CulturePure® is a purified version (≥98% pure) of Cycloheximide that is free of toxic isomers. It is a glutarimide antibiotic and natural fungicide isolated from Streptomyces griseus and a protein synthesis inhibitor in eukaryotic cells. Cycloheximide was discovered by Alma Whiffen-Barksdale of Upjohn Company in 1946 and is now routinely used as a selection agent in several types of isolation media. In molecular biology, it can be used as an experimental tool to determine the half-life of a protein or to study protein synthesis and degradation. Cycloheximide can also be used in protein expression studies, translational profiling and ribosome profiling.
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|Application||Cycloheximide is used in molecular biology for ribosome profiling / translational profiling to understand the complexity of translation initiation. Cycloheximide is used to study protein synthesis, expression and degradation, and determine the half-life of proteins.|
|Mechanism of Action||Cycloheximide binds to the ribosome and inhibits the eEF2-mediated translocation step in protein synthesis, thus blocking translational elongation.|
|Spectrum||Cycloheximide is used for fungi and yeast, including fungi found in brewing test media. It has lower activity against bacteria.|
|Microbiology Applications||Cycoloheximide is routinely used as a selection agent in several types of isolation media: |
Columbia Blood Agar - Campylobacter Selective Supplement (Butzler)
Dermasel Agar - Selective Supplement for dermatophyte fungi
Campylobacter Agar - Campylobacter Selective Supplement (Preston)
Listeria Selective Agar - Listeria Selective Supplement
Listeria Enrichment Broth - Listeria Selective Enrichment Supplement
Listeria Enrichment Broth - Modified Listeria Selective Enrichment Supplement
STAA Agar - STAA Selective Supplement
Legionella CYE Agar - Legionella GVPC Selective Supplement
Campylobacter Agar - Campylobacter Selective Supplement (Karmali)
Bolton Broth - Bolton Broth Selective Supplement
For additional Cycloheximide MIC data, please review the Antimicrobial Index.
|Plant Biology Applications||Cycloheximide is a commonly used lab reagent used in in vitro applications to inhibit fungal growth by targeting protein synthesis. In yeast, concentrations of 200 uM have fungicidal effects (Schneider-Poetsch et al, 2009). The compound can be used as a plant growth regulator to stimulate ethylene production in leaves and fruit.|
|Electrophoresis Applications||Cycloheximide is widely used in biomedical research to inhibit protein synthesis in eukaryotic cells studied in vitro. It inhibits the synthesis of proteins and macromolecules,and affects apoptosis in eukaryotes.|
|Cancer Applications||Pretreatment with Cycloheximide followed by estrogen stimulation prevented the estrogen-induced changes in glucose metabolism in perfused breast cancer T47D clone 11 cells. This suggested that the estrogen stimulation requires synthesis of mRNA and protein (Neeman and Degani, 1989). |
In studying the “immune escape” of cancer cells, in human colorectal cancer cell line COLO 205 is normally resistant to TNF-alpha - a death inducing ligand. However, co-incubation TNF-alpha with Cycloheximide caused time-dependent cell death. In fact, authors found that Cycloheximide sensitizes cells to TNF-alpha-induced apoptosis (Pajak et al, 2005).
|References||Baliga BS, Pronczuk AW and Munro HN (1969) Mechanism of cycloheximide inhibition of protein synthesis in a cell-free system prepared from rat liver. J Biol Chem. 244(16):4480-4489 PMID 5806588 |
Doyle SM, Diamond M and McCabe PF (2010) Chloroplast and reactive oxygen species involvement in apoptotic-like programmed cell death in Arabidopsis suspension cultures. J. Exper. Bot 61 (2):473–482 PMID 19933317
Lee S et al (2012) Global mapping of translation initiation sites in mammalian cells at single-nucleotide resolution. Proc Natl Acad Sci USA. 109(37):E2424-32 PMID 22927429
Neeman M and Degani H (1989) Early estrogen-induced metabolic changes and their inhibition by actinomycin D and cycloheximide in human breast cancer cells: 31P and 13C NMR studies. PNAS 86 (14):5585-5589 PMID 2748604
Pajak B, Gajkowska B, Orzechowski A (2005) Cycloheximide-mediated sensitization to TNF-alpha-induced apoptosis in human colorectal cancer cell line COLO 205; role of FLIP and metabolic inhibitors. J. Physiol. Pharmacol.56 (3)101-118. PMID 16077198
Schneider-Poetsch T et al (2009) Inhibition of eukaryotic translation elongation by cycloheximide and lactimidomycin. Nat. Chem. Biol 6: 209-217 PMID 20118940
Cycloheximide from TOKU-E:
Buchanan BW, Lloyd ME, Engle SM, and Rubenstein EM (2016) Cycloheximide chase analysis of protein degradation in Saccharomyces cerevisiae. J. Vis. Exp. (110), e53975
Cycloheximide Solution from TOKU-E:
Jimenez-Moreno N et al (2019) LIR-dependent LMX1A/LMX1B autophagy crosstalk shapes human midbrain dopaminergic neuronal resilience. bioRxiv 636712 link
|MIC||Candida albicans (CECT 1394)| 12.5 － ?| 408| Mycosphaerella graminicola| 5.62 － 100| 984| Mycosphaerella graminicola (1993 + France)| 68.54 － ?| 984| Mycosphaerella graminicola (1994 + France)| 56.2 － ?| 984| Mycosphaerella graminicola (1995 + France)| 60.04 － ?| 984| Mycosphaerella graminicola (1996 + Germany)| 47.2 － ?| 984| Mycosphaerella graminicola (1997 + Germany)| 62.6 － ?| 984| Mycosphaerella graminicola (1998 + Germany)| 85.4 － ?| 984| Saccharomyces cerevisiae (ATCC 9763)| 0.2 － ?| 812| Saccharomyces cerevisiae (JG436)| 0.05 － ?| 812| Saccharomyces cerevisiae (JGCaMDR1)| 1.6 － ?| 812| Saccharomyces cerevisiae (JGCDR1)| 0.8 － ?| 812| Streptococcus pneumonia (ATCC 6303 + quinolone-susceptible)| >64 － ?| 437| Streptococcus pneumonia (ATCC 7257 + quinolone-resistant)| >64 － ?| 437||