Leptomycin Treatment: Any ideas?? - (Jun/21/2005 )

Hi everyone!
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?
thanks!
sheril

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).

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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).

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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).

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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).

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Ullman K.S. et al, Cell 90 967-970 (1997)

hope that helps