Development of withaferin A analogs as probes of angiogenesis.
The natural product withaferin A (WFA) is a potent angiogenesis inhibitor and it targets the ubiquitin-proteasome pathway in vascular endothelial cells. We generated a biotinylated affinity analog WFA-LC(2)B for use as a probe to study angiogenesis. WFA-LC(2)B inhibits angiogenic sprouting in vitro and it causes levels of ubiquitinated proteins to increase in tumor necrosis factor-alpha-treated human umbilical vein endothelial cells, confirming the retention of WFA's biological activity. We show that WFA-LC(2)B forms protein adducts in endothelial cells which are competed by free WFA in vivo. This WFA-LC(2)B analog will be useful to isolate the biological target of WFA.
Yokota
Yasuno
Y
Department of Ophthalmology and Visual Science, University of Kentucky, Lexington, KY 40536, USA.
Bargagna-Mohan
Paola
P
Ravindranath
Padma P
PP
Kim
Kyung B
KB
Mohan
Royce
R
D020533:Angiogenesis Inhibitors / Q000737:chemistry* / Q000494:pharmacology; D002478:Cells, Cultured; D004591:Electrophoresis, Polyacrylamide Gel; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects; D004875:Ergosterol / Q000031:analogs & derivatives* / Q000737:chemistry / Q000494:pharmacology; D006801:Humans; D015335:Molecular Probes; D054358:Withanolides
D020533:Angiogenesis Inhibitors; D015335:Molecular Probes; D054358:Withanolides; C009684:withaferin A; D004875:Ergosterol