Inhibiting proinflammatory NF-kappaB signaling using cell-penetrating NEMO binding domain peptides.
Nuclear factor kappa B (NF-kappaB) is an inducible transcription factor that regulates the expression of many genes involved in normal immune and inflammatory responses. NF-kappaB activation is normally a rapid and transient response to pro-inflammatory stimuli however dysregulated constitutively active NF-kappaB signaling leads to chronic inflammation and provides a cell survival signal in many types of cancer. NF-kappaB signaling is therefore an important target for the development of novel anti-inflammatory or anti-cancer drugs. We previously identified and characterized a cell-permeable peptide that blocks NF-kappaB signaling by disrupting the critical upstream IkappaB kinase (IKK) complex. We describe in this chapter three separate methods to determine the effects of this NEMO-binding domain (NBD) peptide on pro-inflammatory NF-kappaB signaling in response to tumor necrosis factor (TNF).
Madge
Lisa A
LA
Department of Animal Biology, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA, USA.
May
Michael J
MJ
D002478:Cells, Cultured; D024202:Electrophoretic Mobility Shift Assay; D006367:HeLa Cells / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D051550:I-kappa B Kinase / Q000378:metabolism / Q000494:pharmacology*; D015151:Immunoblotting; D008156:Luciferases / Q000378:metabolism; D016328:NF-kappa B / Q000037:antagonists & inhibitors*; D017434:Protein Structure, Tertiary; D015398:Signal Transduction / Q000187:drug effects*; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
C491731:IKBKG protein, human; D016328:NF-kappa B; D014409:Tumor Necrosis Factor-alpha; D008156:Luciferases; D051550:I-kappa B Kinase