Leptomycin Treatment: Any ideas?? - (Jun/21/2005 )
Has anyone done work with Leptomycin B to block nuclear export of proteins? what concentrations do you normally use it? How much do the cells need to be treated (quantity)? any ideas?
here are some references...
Leptomycin B (LepB) is a potent, specific inhibitor of nuclear export signal (NES)-dependent protein export from the nucleus. Fukuda. M., et al. "CRM1 is responsible for intracellular transport mediated by the nuclear export signal." Nature 390: 308-311 (1997).
The effect of LepB on the CRM1 (exportin 1) NES receptor results from selective alkylation by LepB of a single cysteine residue. Kudo, N., et al. "Leptomycin B inactivates CRM1/exportin 1 by covalent modification at a cysteine residue in the central conserved region." Proc. Natl. Acad. Sci. USA 96: 9112-9117 (1999).
LepB inhibits the nucleo-cytoplasmic translocation of the HIV Rev protein at nanomolar concentrations. Wolff, B., et al. "Leptomycin B is an inhibitor of nuclear export: inhibition of nucleo-cytoplasmic translocation of the human immunodeficiency virus type 1 (HIV-1) Rev protein and Rev-dependent mRNA." Chem Biol. 4: 139-147 (1997).
LepB also exhibits potent antitumor activity in vitro and in vivo, and possesses potent antimycotic and antibiotic activity. Tunac, J.B., et al. "Novel antitumor antibiotics, CI-940 (PD 114,720) and PD 114,721. Taxonomy, fermentation and biological activity." J. Antibiot. 38: 460-465 (1985).
Ullman K.S. et al, Cell 90 967-970 (1997)
hope that helps