Fate of a bioactive fluorescent wortmannin derivative in cells.
Here, we report on NBD-Wm, a fluorescent wortmannin (Wm) probe that maintains the bioactivity of Wm as an inhibitor of PI3 kinase and as an antiproliferative agent. The attachment of the NBD fluorochrome permits NBD-Wm in cells to be monitored by NBD fluorescence-based methods such as FACS or fluorescence microscopy or with an anti-NBD antibody. The fluorescence of NBD-Wm treated cells reached a peak at 1.5 h and then decreased because of the extrusion of a fluorescent compound into the culture media. Cells accumulated NBD-Wm to levels about 30-fold higher than those in the media. NBD-Wm modified five major proteins, with the modification of the catalytic subunit of PI3 kinase being a minor band. The bioactivity of NBD-Wm, coupled with a variety of techniques available for determining its disposition, suggest that NBD-Wm can be a useful tool in understanding the mechanism of action of viridins.
Barnes
Katie R
KR
Center for Molecular Imaging Research, Massachusetts General Hospital and Harvard Medical School, 149, 13th Street, Charlestown, Massachusetts 02129, USA.
Blois
Joseph
J
Smith
Adam
A
Yuan
Hushan
H
Reynolds
Fred
F
Weissleder
Ralph
R
Cantley
Lewis C
LC
Josephson
Lee
L
D000730:Androstadienes / Q000378:metabolism* / Q000494:pharmacology*; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D005456:Fluorescent Dyes / Q000378:metabolism*; D006801:Humans; D042541:Intracellular Space / Q000378:metabolism; D010069:Oxadiazoles / Q000378:metabolism; D000081082:Phosphoinositide-3 Kinase Inhibitors; D013997:Time Factors; D000077191:Wortmannin
D000730:Androstadienes; D005456:Fluorescent Dyes; D010069:Oxadiazoles; D000081082:Phosphoinositide-3 Kinase Inhibitors; C011514:4-nitrobenzofurazan; D000077191:Wortmannin