title: Proteolytic processing of nuclear factor kappa B by calpain in vitro.
abstract: Nuclear factor kappaB (NF-kappaB) is a transcription factor that is critical for the inducible expression of multiple cellular and viral genes. Using the electrophoretic mobility shift assay, we demonstrated that DNA binding activity of NF-kappaB was abolished by proteolysis with mu- and m-calpains in vitro. The proteolysis of NF-kappaB by calpains and hence the abolition of its DNA binding was prevented by calpastatin, calpain inhibitor I and proteasome inhibitor. We also provided evidence that calpains degrade the C-terminal domain of NF-kappaB by Western blot using anti-NF-kappaB (p65) C-terminal antibody. These observations indicate that calpains regulate gene expression through processing of NF-kappaB.
authors.affiliation: Department of Molecular Genetics, Nagoya City University Medical School, Japan.
authors.forename: Z Q
authors.initials: ZQ
authors.lastname: Liu
authors.forename: M
authors.initials: M
authors.lastname: Kunimatsu
authors.forename: J P
authors.initials: JP
authors.lastname: Yang
authors.forename: Y
authors.initials: Y
authors.lastname: Ozaki
authors.forename: M
authors.initials: M
authors.lastname: Sasaki
authors.forename: T
authors.initials: T
authors.lastname: Okamoto
mesh_terms: D001483:Base Sequence; D002135:Calcium-Binding Proteins / Q000494:pharmacology; D002154:Calpain / Q000037:antagonists & inhibitors / Q000378:metabolism*; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D004247:DNA / Q000378:metabolism; D006023:Glycoproteins / Q000494:pharmacology; D006801:Humans; D007976:Leupeptins / Q000494:pharmacology; D008969:Molecular Sequence Data; D016328:NF-kappa B / Q000302:isolation & purification / Q000378:metabolism*; D011499:Protein Processing, Post-Translational
chemical_list: D002135:Calcium-Binding Proteins; D015853:Cysteine Proteinase Inhibitors; D006023:Glycoproteins; D007976:Leupeptins; D016328:NF-kappa B; C039304:calpain inhibitors; C089486:carbobenzoxy-leucyl-leucyl-norvalinal; C033668:calpastatin; D004247:DNA; D002154:Calpain